a) Many bacteria are capable of producing pigments. For example, Serratia marces
ID: 209032 • Letter: A
Question
a) Many bacteria are capable of producing pigments. For example, Serratia marcescens produces a bright red pigment. The purpose of the pigment for this organism is unknown. Your goal is to clone at least one of the genes involved in the production of the pigment in S. marcescens. Your explanation should include the following: explain your rationale for your experimental approach - explain your method for cloning a gene invoved in pigment production as numbered steps - identify the cloning vector that you will use - explain a method for assaying the expressior of this gene that verifies its involvement in the process Tips: Explain the cloning procedure step-by-step. Explain the procedure that you will use for assaying gene expression -don't just name the rrocedure. For example, if PCR is used, identify the primers and expected outcome of the PCR-don't just say "do PCR" as a sterp. Do not explain how the chemistry of PCR or DNA sequencing, but do identify the primers if PCR is used.Explanation / Answer
Objective of the experiment: Cloning and expression of the pswP , gene responsible for color production in S. marcesens
The following steps are required for the cloning of the gene:
(i) The DNA sequence of the pswP, gene is cut with the help of restriction enzyme and inserted into a vector pUC19 vector.
(ii) The pUC19 vector inserted with pswP, gene is transferred into E.coli , known as the process of transformation.
(iii) PCR has been performed with the product made with the template DNA of S. marcescens 274)
(iv) Primer F1 whose sequence is (5'-TTGTAATGCCAAGCTTCTCTCCCGATCGCG-3') and R2 (5'-GGAATTCCGACCGGTACAAAAAGGTCGATT-3') is taken into the study.
(v) Further, electrophoresis is conducted to amplified DNA in order to find mutants.
RT-PCR can be used for estimating the expression of the gene RT-PCR works by measuring the expression of mRNA.
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