The hypothesis to explain the formation of two different mutant phenotypes produced by the transposition of a yeast TE element into the gene for the enzyme X. One mutant lacked complete enzyme activity whereas second mutant have five times more enzyme activity than normal cells. Introduction: Transposable elements in yeast belongs to a family of dispersed and repeated retrotransposons in the genome of Saccharomyces cerevisiae . Saccharomyces cerevisiae is an important model organism to study the LTR (long terminal repeats)-retrotransposons biology. Five retrotransposon families are present in S. cerevisiae, designated as Ty1-Ty5. These Ty elements have a tendency to influence the genome organization. These Ty elements are the causal agents of mutation and genome rearrangment through recombination.

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Clary Fray used the pET vector system to express her prokaryotic amylase enzyme. She added peptone into her culture broth of BL21(DE3) Escherichia coli strain to induce protein expression. At the end of the experiment, she discovered that her protein was not expressed. She repeated three more times but her protein of interest was still not produced. (i) (ii) (iii) (iv) (v) Explain the reason why Clary failed to obtain her protein of interest and suggest a solution to troubleshoot this problem. Clary plans to express her protein along with a polyhistidine-tag. Explain the importance of His-tag in protein work. Is DH5a Escherichia coli suitable to propagate the plasmid before protein expression? Besides heat shock method, elaborate another procedure Clary could utilize to transform the recombinant pET vector into the host cell. If her supervisor instructs her to express a gene from gold fish (Carassius auratus), is the expression system she is using now suitable for this experiment?…

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Chapter 15, Problem 15TYK

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The hypothesis to explain the formation of two different mutant phenotypes produced by the transposition of a yeast TE element into the gene for the enzyme X. One mutant lacked complete enzyme activity whereas second mutant have five times more enzyme activity than normal cells.

Introduction:

Transposable elements in yeast belongs to a family of dispersed and repeated retrotransposons in the genome of Saccharomyces cerevisiae. Saccharomyces cerevisiae is an important model organism to study the LTR (long terminal repeats)-retrotransposons biology. Five retrotransposon families are present in S. cerevisiae, designated as Ty1-Ty5. These Ty elements have a tendency to influence the genome organization. These Ty elements are the causal agents of mutation and genome rearrangment through recombination.

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Clary Fray used the pET vector system to express her prokaryotic amylase enzyme. She added peptone into her culture broth of BL21(DE3) Escherichia coli strain to induce protein expression. At the end of the experiment, she discovered that her protein was not expressed. She repeated three more times but her protein of interest was still not produced. (i) (ii) (iii) (iv) (v) Explain the reason why Clary failed to obtain her protein of interest and suggest a solution to troubleshoot this problem. Clary plans to express her protein along with a polyhistidine-tag. Explain the importance of His-tag in protein work. Is DH5a Escherichia coli suitable to propagate the plasmid before protein expression? Besides heat shock method, elaborate another procedure Clary could utilize to transform the recombinant pET vector into the host cell. If her supervisor instructs her to express a gene from gold fish (Carassius auratus), is the expression system she is using now suitable for this experiment?…

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