Cloning and phylogenetic analysis of RNA helicase p68 encoding DDx5 gene from human MCF7 cell line
2016
Shaheen, S. (University of Agriculture, Faisalabad (Pakistan). Dept. of Chemistry and Biochemistry) | Jamil, A. (University of Agriculture, Faisalabad (Pakistan). Dept. of Chemistry and Biochemistry) | Iqbal, T. (University of Agriculture, Faisalabad (Pakistan). Dept. of Chemistry and Biochemistry) | Ashraf, M. (University of Agriculture, Faisalabad (Pakistan). Dept. of Botany)
In this study, genomic DNA was isolated from MCF7 cells and used as template for polymerase chain reaction (PCR) to amplify p68 by using sequence specific primers. The PCR product was purified, ligated into pMSCV-GFP vector and transformed into E. coli DH5 alpha competent cells. Restriction analysis was performed to confirm the cloning. Further, the gene was sequenced, and on the basis of this partial sequence, a phylogenetic tree was made to access homology among different organisms. In present study, gene encoding p68 in human was found to be more close to gene encoding p68 in Sus scrofa and Pan troglodytes. The results obtained from present study will be helpful to study the role of p68 in regulation of gene expression in humans in future.
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