In bacteria of the genus Salmonella, the hilD gene encodes a transcriptional reg

Question

In bacteria of the genus Salmonella, the hilD gene encodes a transcriptional regulator, HilD. This regulator is responsible for integrating a large number of signals which allows the cell to express a type three secretion system at a time appropriate to allow it to invade the host's intestinal epithelium. You've been tasked with analyzing HilD. In order to produce enough of the protein, it will be necessary to ctone the gene into an expression vector. The first step of this process will be to amplify the hilD gene via PCR. 1. Given the sequence on the following page, choose forward and reverse primers that will amplify the underlined portion of the hilD gene. a. Underline the primer sequence on the following page. Then write out your primers below and indicate the 5' and 3' ends. Remember that the 3' or reverse primer is the reverse complement of the template (think about which direction DNA extends). (5 points) b. What is the size of your target DNA? (Note: each line contains 70 nucleotide bases) (5 points)

Explanation / Answer

Q.1

A.)

Forward Primer: 5' - ATG GAA AAT GTA ACC TTT - 3'

It is the same as template strand running from 5' to 3' direction.

Reverse Primer: 3' - TTT TAC CGC TTG GTA ATT - 5'

It is the reverse complement of 5' - AAA ATG GCG AAC CAT TAA - 3'

B.)

Total DNA Content:

33 LINES *70 = 2310

2310 + 14 (Last Line) = 2324 bp.

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Target DNA - 930 bp.

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