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Question
Commonly used reporter gene that utilizes histo-chemical assay in plant expression vector is
Commonly used reporter gene that utilizes histo-chemical assay in plant expression vector is
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Solution
The correct option is A CAT
Genes whose phenotype can be discerned by visual examination of colonies growing on a plate and/or ones that can be used to measure levels of gene expression are reporter genes. Chloramphenicol acetyltransferase or CAT which transfers acetyl groups to chloramphenicol and thus, rendering it harmless. The promoter of interest is placed upstream of the CAT coding sequence to confirm its activation and either mRNA or protein production is measured by the histo-chemical assay. The extracted protein from the cells is mixed with chloramphenicol having radioactive hydrogens and acetyl-CoA to measure the amount of acetylated chloramphenicol and thereby amount of protein synthesized due to activated promoters. lac Z gene encodes β-galactosidase and presence or absence of β-galactosidase activity is easily detected by growing cells on medium containing the chromogenic substrate X-gal. The gusA gene encodes β-glucuronidase which cleaves 4-methylumbelliferyl-b-D-glucuronide (MUG) to release a blue pigment. Both lacZ and gusA genes produce the visible phenotype that does not need any enzyme assay as is the case with CAT gene. The correct answer is C.
Genes whose phenotype can be discerned by visual examination of colonies growing on a plate and/or ones that can be used to measure levels of gene expression are reporter genes.
Chloramphenicol acetyltransferase or CAT which transfers acetyl groups to chloramphenicol and thus, rendering it harmless. The promoter of interest is placed upstream of the CAT coding sequence to confirm its activation and either mRNA or protein production is measured by the histo-chemical assay.
The extracted protein from the cells is mixed with chloramphenicol having radioactive hydrogens and acetyl-CoA to measure the amount of acetylated chloramphenicol and thereby amount of protein synthesized due to activated promoters. lac Z gene encodes β-galactosidase and presence or absence of β-galactosidase activity is easily detected by growing cells on medium containing the chromogenic substrate X-gal. The gusA gene encodes β-glucuronidase which cleaves 4-methylumbelliferyl-b-D-glucuronide (MUG) to release a blue pigment. Both lacZ and gusA genes produce the visible phenotype that does not need any enzyme assay as is the case with CAT gene. The correct answer is C.
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