Question
please I need help with question 5 and 6 buy you can choose to answer just any of them if both are too much for you.Thanks
Question 1 The information listed below summarizes the current state of our knowledge concerning Syndrome the data presented are real The actual name of has been fabricated so as not to deprive our students of the the syndrome to formulate responses to the questions that follow the intellectual joy derived from utilizing the data summary given below Syndrome X is a man genetic disorder that results from a defect at one genetic locus and patients with the disorder manifest ple of the anterior of that occur during embryonic development These effects include Syndrome X patients), mild the eye (which causes the eye disease known as glaucoma in used to head and face malforma ns and underdeveloped teeth. One of the pedigrees determine the mode of inheritance of the disorder is found below b) Families displaying the disease were examined for linkage of the disease locus to a polymorphic site unique to each family. From such studies scientists were able to report linkage of the gene that is defective in the disorder to 4. (From this point on we will use the term o designate the normal form of the gene that is defective in Syndrome X) c) Investigators used positional clo gene when one knows nothing ng a methodology for isolating a cDNA clone co screening a cDNA about the function of the protein encoded by the gene and has no probe for Subsequently, library, and ultimately isolated a fu ength cDNA clone coding for the normal gene they isolated a human genomic clone coding for gene x d) The cDNA clone as well as parts of the genomic clone were sequenced. The results are shown in Fig. 1 e) since nothing was known about the structure or function of the protein encoded by gene x, scientists deduced its amino acid sequence from the cDNA nucleotide sequence of gene x and it studies. The deduced amino acid sequence was analyzed via software capable of predicting with some degree of accuracy, the secondary structure of a protein (but not the complete tertiary structure and presence of a helix-turn-helix was revealed the f) Mutant gene x isolated from members of 10 different families diagnosed with Syndrome X were analyzed to identify the mutations the changed y showed the following Nucleotide num ber 981 was from a G to an A as shown in Pig 1. The other nine families showed other distinct mutations g) It is often helpful when studying human genetic disorders to have a mouse model of the discase as well as a cDNA clone for the gen deduced amino acid sequence under study. For this reason, scientists isolated a mouse cDNA clone whose showed a 91% similarity to the deduced amino acid sequence for human gene Utilizing all of the data presented, in conjunction with the many concepts that you have learned this semester, answer th questions that follow.
Explanation / Answer
5) cDNA library preparation
isolate mature mRNA from the brain tissue of the mouse. using reverse transcriptase enzyme convert the mRNA to cDNA. ligate the cDNA using DNA ligase with a plasmid after restriction enzyme digestion of the plasmid. transform the plamid to bacterial cells. use lambda phage vectors for cloning
