Answer to Question #350068 in Genetics for aleace

Question

A genetics research group has determined the sequence of a rat gene thought to be

involved in controlling the fertilising ability of rat sperm. They believe a similar gene may

control fertility in humans. Describe how what is known about the rat gene can be applied

in PCR to clone the complementary human gene. You should explain the experimental

approach taken and how it could confirm that the human gene in question does indeed

correspond to the rat gene.

Answer 1

This gene is called SPAG11 (SPerm associated AntiGen 11), which controls fertility in both rats and humans by promoting sperm maturation. These genes are the orthologs in rats and humans so that rat SPAG11 can be used to clone human SPAG11.

According to the article (available here: https://rbej.biomedcentral.com/articles/10.1186/1477-7827-4-23), when performing PCR, cDNAs obtained from one of the transcripts (Spag11c or Spag11t) from the rat Rattus norvegicus are used with application of the following primers:

• Spag11c: forward – 5' CTG CAG TCC CCT CCA CAG CC 3'; reverse – 5' GTG TGC AGG TCA CTT CAA CTT C 3';
• Spag11t: forward – 5' CTG CAG TCC CCT CCA CAG CC 3'; reverse – 5' CAT CCA CGC TGT CAC CTC CC 3'.

The amplified cDNA can be inserted into any eukaryotic vector and cloned in cultures of human testicular cells that have a mutation in the SPAG11 gene. The result of the effectiveness of the above technique will be that human testicular cells will acquire a functional analogue of the SPAG11 gene derived from rat and will develop into testicular tissues in which human spermatozoa will mature.