A) Name the two (2) sequence elements that must be in the plasmid at position X

Question

A) Name the two (2) sequence elements that must be in the plasmid at position X for the gene to be transcribed and translated. Briefly describe the function of each sequence element.

B) The host cells also need two additional sequence elements to maintain and copy the plasmid. These are shown as ‘Sequence 1’ and ‘Sequence 2’. What are the correct names of these two (2) sequence elements? Briefly describe the function of each sequence element.

C) Design a forward PCR primer that you could use as part of a forward and reverse primer pair to amplify the entire TINY gene. Write the primer in your answer book, indicating the 5 and 3 ends of the sequence. Show your working for the primer design.

D) Assume the first three bases of the TINY gene sequence shown are the start codon (ATG) and the last three bases are a stop codon (TAG). How many amino acid residues are in the encoded protein? Explain your answer briefly.

E) What is the sequence of the first ten bases of the mRNA produced from the TINY coding sequence?

Q1. The following is the DNA sequence of a hypothetical gene for the TINY protein. It is called the TINY gene. 1 atgatcatca actgt cacga ccaattcatt gtatgaaaag 41 acacagcaag caggatatag Imagine you are doing a research project in a laboratory and your supervisor asks you to clone the TINY gene into a plasmid vector so you can express the gene in an E. coli bacterial host. The process you use is shown below. TINY gene TINY Plasmid Vector Sequence 1 Sequence 1. Sequence 2 Sequence 2 The direction of TINY gene transcription is shown by the dotted arrow. Other sequences that are necessary in the plasmid are shown with an X, and with circles labelled 'Sequence 1' and 'Sequence 2'

Explanation / Answer

1. Promoter region and restriction site. Promoter region is the region at which the RNA polymerase binds and initiates the process of transcription while the restriction sites are speciic patterns of nucleotides that are recognized by the restriction enzymes which functions in the cleavage of the vector molceule so that the gene of interest can be inserted in its place.

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