6.2: Blood Type Genetics Teacher's Preparation Notes

Overview

In this minds-on, hands-on activity, students will learn the genetics and immunobiology of the ABO blood type system. Students will use simple chemicals to simulate blood type tests and then carry out genetic analyses to determine whether hospital staff accidentally switched two babies born on the same day. This activity reinforces students' understanding that genes code for proteins which influence an organism’s characteristics and Punnett squares summarize how meiosis and fertilization result in inheritance. Students will also learn the concept of codominance.

Optional additions for the Student Handout is presented in the last two pages of these Teacher Preparation Notes. Students analyze the genetics of skin color to understand how fraternal twins could have very different skin colors. In this optional addition, students learn the concept of incomplete dominance, the difference between incomplete dominance and codominance, and how multiple genes and the environment can influence a single phenotypic characteristic.

As background for this activity, students should have a basic understanding of:

• Dominant and recessive alleles, with heterozygous individuals having the same phenotype as homozygous dominant individuals
• How meiosis and fertilization result in inheritance and how these processes are summarized in Punnett squares.

To provide this background you may want to use the first three pages of our "Genetics" activity or the first four pages of "Genetics Supplement" (both available at http://seren dip.brynmawr.edu/sci_edu/waldron/#genetics).

Learning Goals Related to National Standards

In accord with the Next Generation Science Standards and A Framework for K-12 Science Education:

• Students will gain an understanding of several Disciplinary Core Ideas:
  • LS1.A: Structure and Function –"All cells contain genetic information in the form of DNA molecules. Genes are regions in the DNA that contain the instructions that code for the formation of proteins."
  • LS3.A: Inheritance of Traits – "The instructions for forming species' characteristics are carried in DNA."
  • LS3.B: Variation of Traits – In sexual reproduction, meiosis can create new genetic combinations and thus more genetic variation.
• Students will engage in several Scientific Practices:
  • Constructing explanations
  • Engaging in argument from evidence
  • Carrying out an investigation
  • Analyzing and interpreting data.
• This activity provides the opportunity to discuss the Crosscutting Concept, "Structure and Function".
• This activity helps to prepare students for the Performance Expectations
  • HS-LS3-1, "Ask questions to clarify relationships about the role of DNA and chromosomes in coding the instructions for characteristic traits passed from parents to offspring."
  • HS-LS3-2, "Make and defend a claim based on evidence that inheritable genetic variations may result from: (1) new genetic combinations through meiosis…"

 

Specific Learning Goals

• There are four blood types in the ABO system: A, B, AB, and O. These blood types refer to the presence or absence of two different versions of a carbohydrate molecule (A and B) on the surface of red blood cells.
• Genes code for proteins which influence a person's characteristics. The ABO blood type gene codes for a protein enzyme that can attach carbohydrates to the surface of red blood cells. This gene has three alleles: the I^A allele codes for a version of the enzyme that attaches the A carbohydrate; the I^B allele codes for a version of the enzyme that attaches the B carbohydrate; and the i allele codes for an inactive protein that does not attach either carbohydrate.
• As a result of meiosis and fertilization, each person inherits one allele of this gene from his/her mother and a second allele from his/her father. The results of meiosis and fertilization are summarized in Punnett squares.
• Both inherited alleles code for the production of proteins in red blood cell precursors. In a heterozygous person with the I^Ai or I^B i genotype, the single copy of the I^A or I^B allele in each cell codes for enough enzyme to result in type A or type B blood, respectively. Thus, the i allele is recessive relative to the I^A or I^B alleles.
• Codominance refers to inheritance in which two alleles of a gene each have a different observable effect on the phenotype of the heterozygous individual. The I^A and I^B alleles are codominant since a person who has the I^AI^B genotype has type AB blood. Both the I^A and I^B alleles are active so the cells produce both the version of the enzyme that attaches the A carbohydrate to the surface of red blood cells and the version of the enzyme that attaches the B carbohydrate, resulting in type AB blood.
• Both the A and B carbohydrates are antigens which stimulate the formation of antibodies. Antibodies are special proteins that travel in the blood and react with specific antigens. For example, anti-A antibodies react specifically with A antigens on the surface of red blood cells, and anti-B antibodies react specifically with B antigens.
• Normally, your body does not make antibodies against antigens which are part of your own body. For example, a person with type A blood does not make anti-A antibodies but does make anti-B antibodies. A blood transfusion can harm a person if the donated red blood cells have antigens that react with antibodies in the person's blood.

Supplies, Preparation, and Suggestions for Implementation

Supplies

(see pages 3-4 for information about amounts needed)

• Synthetic blood of all four blood types (A, B, AB, and O)
• Solution with synthetic anti-A and anti-B antibodies
• Drop-controlled bottles or small bottles, each with a dropper or pipette (can be reused in multiple classes; if contamination occurs, you may need to wash and refill the bottles between classes)
• Small non-porous testing surfaces suitable for mixing blood and antibodies, e.g. blood-typing trays, microscope slides or white plastic lids (can be washed and reused in multiple classes) (Unless you are using the pre-labeled blood-typing trays, you will want to use a marker to identify two different spots where students will test for the type A antigen and for the type B antigen. You will also need some way for students to keep track of whose blood is on which testing surface.)
• Toothpicks for mixing blood and antibody solution (Each toothpick should be disposed of immediately after both ends have been used.)
• Containers such as soda or water bottles to use as trash containers so the students can throw away their toothpicks immediately after use to avoid contamination

Implementation If You are Using Purchased Synthetic Blood and Antibodies

To determine the amount of supplies you will need, you should choose among these three recommendations for implementation (or decide on your own approach).

• Give each student group at their lab table:
  • 7 bottles with the blood samples for each subject listed in the table on the bottom of page 4 of the Student Handout
  • 1 bottle with the anti-A antibody solution and another bottle with the anti-B antibody solution
  • 7 testing surfaces, e.g. microscope slides or white plastic lids
  • 7 toothpicks (if you have the students use both ends of each toothpick); otherwise 14 toothpicks.
• Alternatively, you can use seven bottles with blood samples, a bottle with the anti-A solution, and a bottle with the anti-B solution to set up three stations, one where each student group will get the blood samples for each subject, one where they will get the drops of anti-A antibody solution, and one where the they will get the drops of anti-B antibody solution. Each student group will need their own set of seven testing services and toothpicks.
• If you want to minimize supplies and experimental time, you can set up the three stations as described in the previous paragraph, but have each student group test the blood sample from a different subject and then combine the results from the different student groups to complete the table on the bottom of page 4 of the Student Handout. For this approach, you will need to make minor modifications in the Procedure instructions on page 4 of the Student Handout.

If you use either of the first two recommended approaches described above, each student group will carry out seven blood type tests. For each blood type test, you will need two drops of anti-A antibody solution and two drops of anti-B antibody solution and four drops of blood (see Preparation section below for suggested blood types for each subject). This amounts to 14 drops of each kind of antibody solution and 28 drops of blood for each student group. There are approximately 15-20 drops in each milliliter of solution, so you will need approximately 1 mL of each type of antibody solution for each student group (although you will probably want more to be prepared for student error such as using too many drops or contamination). The amounts of blood of each type needed will vary, depending on your choice of blood types for each subject (see Preparation section below). If you use the third recommended approach above, each student group will carry out one blood type test so you will need substantially less of each type of antibody solution and blood.

You can purchase synthetic blood (type A, type B, type AB, and type O) and synthetic anti-A and anti-B antibody solutions in 500 mL bottles (from Frey Scientific and CBO Science for a total of $95 for all six items as of mid-2016; https://store.schoolspecialty.com/OA_HTML/xxssi_ibeSearchResults.jsp?resetSearch=true&type=search&searchType =productResults&minisite=10029&query=blood+typing+serum&idx=&relevancy=&ps=&r=&refQuery=&requiredFields=&searchType=&minisite=10029). If you want, you can also purchase blood-typing trays (https://store.schoolspecialty.com/OA_HTML/ibeCCtpItmDspRte.jsp?minisite=10029&item=51677).

If you are doing the activity with only one or two classes, it may be more economical to purchase kits (and/or refills) from

• Carolina for $43 (and/or $24); http://www.carolina.com/blood-typing/carolina-abo-rh-blood-typing-with-synthetic-blood-kit/FAM_700101.pr?question=700101. We recommend you not use the Rh antiserum included in this kit.)
• Ward’s Science for $48 (and/or $29); https://www.wardsci.com/store/catalog/product.jsp?catalog_number=366809.

These kits have additional supplies such as some dropper bottles and testing trays. You will probably want to contact these companies to verify that their kits have the blood types and quantities you will need.

A Cheaper Alternative

If you have insufficient budget for these commercial products, you can use the following economical alternative. You can make simulated blood by combining 0.25 L of milk with red food coloring until the solution is bright red, and then adding a drop of green food coloring for a dark red color. You will need to give your students different anti-A and anti-B simulated sera, depending on what type blood the sample is supposed to contain.

Type of Blood	Simulated Anti-A Solution Contains:	Simulated Anti-B Solution Contains:
A	White vinegar	Water
B	Water	White vinegar
AB	White vinegar	White vinegar
O	Water	Water

To avoid confusion, you will probably want to have each student group test the blood type of only one of the subjects. Also, if you use this approach, you should change the instructions on page 4 of the Student Handout to direct students to use three drops of the blood and three drops of each type of antibody solution for each sample.

Preparation

Before class, you should prepare a bottle with the appropriate blood sample for each person listed in the table on page 4 of the Student Handout and label the bottle with the person’s name. You may want to vary the blood types in these samples for different classes, in order to maintain some variety and suspense. The table on the next page illustrates some possible combinations of blood types for each person. You can make other combinations, provided that:

• Michael Jr. can be the son of Michael and Danielle
• One of the baby girls could be a daughter of one of the couples and could not be a daughter of the other couple. The other baby girl could be a daughter of the other couple.

Examples of Blood Type Combinations You Can Use

	1	2	3	4	5	6	7	8	9	10
Michael (father of twins)	AB	AB	AB	AB	O	A	B	AB	A	A
Danielle (mother of twins)	O	A	B	AB	AB	AB	AB	AB	O	A
Earnest (father of daughter)	A	A	A	A	B	B	B	B	A	A
Denise (mother of daughter)	B	B	B	B	A	A	A	A	B	B
Michael Jr. (boy twin)	A	A	A	A	B	B	B	B	A	A
Baby girl 1 (girl twin, according to hospital)	B	B	B	B	A	A	A	A	O	O
Baby girl 2 (daughter of Earnest and Denise, according to hospital)	O	O	O	O	O	O	O	O	B	B

Each column of this table will also work if you reverse the blood types for the two baby girls. This will add suspense about whether the hospital made a mistake. However, if the hospital made a mistake and the twins have similar skin color, and if you are using the Optional Addition to the Student Handout (provided on the last two pages of these Teacher Preparation Notes), you will need to change some of the wording on the first page of the Optional Addition.

General Instructional Suggestions

To maximize student learning, we recommend that you have your students complete groups of related questions in the Student Handout, individually or in pairs, and then have a class discussion for each group of related questions. In each discussion, you can probe student thinking and help them to develop a sound understanding of the concepts and information covered before moving on to the next part of the activity.

The PDF of the Student Handout shows the correct format; please check this if you use the Word document to make revisions.

If you would like to have a key with the answers to the questions in the Student Handout, please send a message to [e-mail]. The following paragraphs provide additional instructional suggestions and background information – some for inclusion in your class discussions and some to provide you with the relevant background that may be useful for your understanding and/or for responding to student questions.

 

Biology Background and Suggestions for Discussion

For the ABO blood group:

• The I^A allele codes for a version of an enzyme that plays a crucial role in synthesizing glycoprotein and glycolipid molecules that contain the Type A carbohydrate; these glycoproteins and glycolipids are located in the cell membrane of red blood cells.
• The I^B allele codes for a different version of this enzyme that plays a crucial role in synthesizing glycoproteins and glycolipids with the Type B carbohydrate molecules.
• The i allele codes for an inactive version of the enzyme.

The function of these carbohydrate molecules is unknown. In general, people who have Type O blood with neither Type A nor Type B carbohydrates are as healthy as people who have the Type A and/or Type B carbohydrates. Different blood types are correlated with certain illnesses and vary in frequency in different ethnic groups, but the reasons are unknown.

In discussing question 2, you should remind students that heterozygous individuals have the same phenotype as an individual who is homozygous for the dominant allele. You will probably also want to point out that recessive alleles often code for a nonfunctional protein. In a heterozygous individual, a single dominant allele can code for enough functional protein to result in the same phenotype as the phenotype of the homozygous dominant individual. For example, the i allele is recessive relative to the I^A or I^B alleles because, in a heterozygous individual, the single dominant I^A or I^B allele codes for enough functional enzyme to result in the same blood type as observed in a homozygous dominant individual.

This activity helps students to understand the molecular basis for codominance, as well as dominant-recessive alleles. Each cell in the body contains two copies of each gene and typically both alleles are transcribed. Thus, at a molecular level, the alleles of most genes are codominant. For example, the I^A I^B genotype results in the production of both the version of the enzyme that puts Type A carbohydrate molecules on red blood cells and the version of the enzyme that puts Type B carbohydrate molecules on red blood cells. Therefore, the I^A I^B genotype results in Type AB blood. This illustrates codominance at the phenotypic level.

In discussing questions 5 and 9, you will want to point out how meiosis and fertilization result in new combinations of alleles, so children may have different blood types and other phenotypic characteristics than their parents have. Of course, it also should be pointed out that the transmission of genes via meiosis and fertilization result in similarities between offspring and parents. For example, a child can only have type A or AB blood if one or both parents have type A or AB blood (i.e. a child with the I^A allele must have at least one parent with this allele).

In discussing the section on “Understanding Blood Type Tests”, you may want to point out that each antibody has two sites that bind to antigens, as shown in the figures on page 3 of the Student Handout.

You may want to mention that some types of antibody bind to the antigens on the surface of bacteria that have infected a person’s body. This figure shows one way that antibodies can contribute to the destruction of bacteria and thus protect our bodies against infection. A macrophage is a phagocytic cell that can ingest a bacterium, kill it, and then digest it.

Normally, your body does not make antibodies against any molecules that are part of your own body. This is useful because antibodies against antigens that are part of your body could trigger harmful reactions, such as an immune attack on your body cells. As would be expected, a person does not make antibodies against the blood type antigens present on their red blood cells. However, a person with type A blood does make anti-B antibodies since gut bacteria have antigens similar to type B antigens, and this stimulates the production of anti-B antibodies. If a person with type A blood is given a transfusion of type B blood, the anti-B antibodies will cause the donated type A red blood cells to clump in a transfusion reaction that can block blood vessels and even cause death. Similarly, a person with type B blood will have a transfusion reaction if given type A blood.

The ABO blood types are the major determinant of which type of blood will cause a transfusion reaction. However, the determination of blood type is more complex than the ABO blood types. For additional information on other blood group antigens and blood types, see http://www.ncbi.nlm.nih.gov/books/NBK2264/.

On page 5 of the Student Handout, students use genetic analysis of the blood type results to determine whether the babies were switched. Modern methods use DNA testing to determine biological relatedness; these results are much more definitive than testing blood types (http://en.Wikipedia.org/wiki/Parental_testing).

 

Optional Addition to Student Handout

This Optional Addition to the Student Handout is shown on the last two pages of these Teacher Preparation Notes. This analysis of the genetics of skin color introduces students to:

• The concept of incomplete dominance
• The difference between codominance vs. incomplete dominance
• The influence of multiple genes and environmental factors on a single phenotypic characteristic.

Skin color is influenced by multiple genes. For example, one gene that influences skin color codes for the enzyme tyrosinase, a crucial enzyme involved in the synthesis of melanin, the primary pigment in skin and hair. The normal allele codes for functional tyrosinase and the allele for albinism codes for a defective version of this enzyme. The allele for albinism is recessive because, even when there is only one copy of the normal allele, this allele codes for enough functioning enzyme to produce enough melanin to result in normal skin and hair color.

Another important gene that influences skin color is the MC1R gene which codes for the melanocortin receptor; when the alpha-melanocyte-stimulating hormone binds to normal melanocortin receptor this stimulates melanocytes to produce melanin. More than 80 alleles of the MC1R gene have been identified, resulting in various levels of function of the melanocortin receptor and correspondingly varied skin tones. Heterozygotes for these alleles have intermediate skin color, between the lighter and darker homozygotes (called incomplete dominance or a dosage effect). The multiple alleles and the effects of incomplete dominance result in multiple different phenotypes for skin color (and hair color). (Additional information on this gene is available at https://ghr.nlm.nih.gov/gene/MC1R. Additional information on the complex genetics and molecular biology involved in the regulation of skin color is available at http://www.jbc.org/content/282/38/27557.full and http://hmg.oxfordjournals.org/content/18/R1/R9.full.)

In discussing question 11, the following table may be helpful.

Type of Dominance	Phenotype of Heterozygous Individual
Dominant-recessive pair of alleles	Same as the phenotype of individual who is homozygous for the dominant allele
Codominance	Shows different observable phenotypic effects of both alleles; phenotype different from either homozygous individual
Incomplete dominance	Intermediate between phenotypes of the two types of homozygous individual (typically observed for quantitative traits); phenotype different from either homozygous individual

The Optional Addition to the Student Handout provides the opportunity to reinforce student understanding that individual phenotypic characteristics are often influenced by multiple alleles of multiple genes, as well as environmental factors. Our introductory genetics teaching frequently focuses on inheritance and phenotypic effects of single genes, as illustrated by the first page of the Optional Addition. However, this is only a beginning for understanding the genetics of most traits. For example, as discussed on the second page of the Optional Addition, a person with a Bb genotype could have lighter or darker skin, depending on whether he or she:

• Has developed a tan as a result of sun exposure or tanning booth use
• Has alleles for other genes that contribute to darker skin color.

During your discussion of question 13, you may want to explain that the genotype/phenotype table on the preceding page of the Student Handout is a simplified introduction to the genetics of skin color. The figure below provides a somewhat more accurate representation. Even this relatively complex Punnett square is a simplified representation of reality since it assumes a simple additive model with only two alleles and incomplete dominance for each of the three genes shown.

Additional Activities

"Genetics – Major Concepts and Learning Activities" (http://serendip.brynmawr.edu/exchange/bioactivities/GeneticsConcepts)

This overview summarizes important genetic concepts and proposes an integrated sequence of learning activities to develop student understanding of these key concepts. Part I provides an outline of key concepts needed to understand how genes influence phenotypic characteristics and how genes are transmitted from parents to offspring. Part II recommends an integrated sequence of learning activities to develop student understanding of these key concepts. These learning activities are aligned with the Next Generation Science Standards. Part III suggests supplementary and alternative learning activities.

Optional Addition to Student Handout

Why Do the Twins Look so Different?

Now, Danielle wants to know how her twins could look so different, with Michelle having light skin and Michael Jr. having dark skin. First, Danielle needs to understand that there are two types of twins. Identical twins have exactly the same genes since identical twins originate when a developing embryo splits into two embryos.

10. How do you know that Michelle and Michael Jr. are not identical twins?

 

Michelle and Michael Jr. are fraternal twins, the result of two different eggs, each fertilized by a different sperm. These different eggs and sperm had different alleles of the genes for skin color. Therefore, Michelle and Michael Jr. inherited different alleles of these genes, so they have different skin colors.

To begin to understand how Michelle could have light skin and her twin brother, Michael Jr., could have dark skin, we will consider two alleles of one of the genes for skin color. Notice that, for this gene, a heterozygous individual has an intermediate phenotype, halfway between the two homozygous individuals.

Genotype	Phenotype (skin color)
BB	Dark Brown
Bb	Light Brown
bb	Tan

When the phenotype of a heterozygous individual is intermediate between the phenotypes of the two different types of homozygous individual, this is called incomplete dominance.

11a. Explain how incomplete dominance differs from a dominant-recessive pair of alleles. (Hint: Think about the phenotypes of heterozygous individuals.)

 

11b. Explain how incomplete dominance differs from co-dominance.

12. The parents, Michael and Danielle, both have light brown skin and the Bb genotype. Draw a Punnett square and explain how these parents could have two babies with different color skin – one dark brown and the other tan.

Obviously, people have many different skin colors, not just dark brown, light brown, or tan. One reason for the many different skin colors is that skin color is influenced by multiple genes with multiple alleles. Scientists have found that:

• Different skin colors result from differences in the types and amounts of the pigment melanin in skin cells.
• Several different proteins influence the production and processing of melanin molecules in skin cells.
• Different alleles of the genes that code for these proteins result in different skin colors.

Environmental influences also affect skin color. For example, exposure to sunlight can change the activity of genes that influence skin color and increase the amount of melanin in skin cells.

 

This flowchart summarizes the multiple genetic and environmental influences on skin color.

13. This information indicates that the chart on the previous page is oversimplified. Multiple factors influence skin color, so two people who both have the Bb genotype can have different skin colors. For example, Hernando and Leo both have the Bb genotype, but Hernando’s skin is darker than Leo’s. Explain two possible reasons why Hernando and Leo have different skin colors.