Now you want to study the protein product of this mutant brat gene, and you want...

Now you want to study the protein product of this mutant brat gene, and you want to introduce this human brat gene into a bacterial expression vector (one that can be expressed to make a protein in bacterial cells) in hopes of producing a large amount of the human mutant Brat protein. Should you use the genomic DNA or the cDNA version of this gene? Explain your reasoning.

Expert Solution

We should definetely use cDNA version of brat gene.

The reason behind it is, difference in organization of genes in bacteria (a prokaryote) and human (an eukaryote). Genes in prokaryotes are contiguous i.e. they are arranged one after another without any gap. Whereas in case of eukaryotes, discontinuous segments of genes are found. The coding segments of DNA (Exon) are broken by presence of non coding DNA (introns). These non coding introns are removed after transcription of DNA into mRNA. This process of intronic removal is called Splicing. The spliceosome which does the splicing is not found in prokaryotes or bacteria as there is no need of splicing.

So, if We will introduce DNA in the bacteria, then it will contain both exon and intron. Bacteria would not be able to remove intron from transcribed mRNA because of lack of such machinary. Hence, proper protein will not be synthesized by the ribosome.

On the contrary when we will introduce cDNA which is made from mRNA by the enzyme reverse transcriptase. Then it will not contain any intron (because it is reverse transcribed from already spliced mRNA ). Bacteria will easily translate such mRNA transcibed from cDNA. Hence we will get proper mutant brat protein.

gladiator answered 2 years ago

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