Sequence Analysis,Cloning and Induced Expression of Chaperonin Gene in Housefly（Musca domesitca）

Abstract

Abstract: The aim of this study is to analyze and predict the structural and characteristics of genes and encoding proteins of MD-TCP Ⅰ（Musca domesitca Chaperonin TCP- 1（MD-TCPⅠ））, with the methods of cloning and expressing that gene. Sequence analysis indicated that the open reading frame was 753 bp, encoding a putative protein consisting of 250-amino acids, which no signal sequence and NCBI-BLAST showed acid sequence identify with other insect TCP-1 were 89%. The protein, with a predicted molecular weight of 27.07 kD, and pI of 5.92, which acid sequence as tcp-1 belong to HSP60 family. The gene coding for MD-TCP Ⅰwas amplified by polymerase chain reaction（PCR）, and then was ligated into vector pET-28a（+）and transformed into Escherichia coli BL21（DE3）competent cell, induced with IPTG. The fusion protein in the expression vector was analyzed by SDA-PAGE. The results indicated that the recombinant plasmid with the correct target gene was constructed, and the fusion protein was expressed in E. coli BL21（DE3）.

Key words: Musca domestic, TCP, Sequence analysis, Gene cloning, Express

Zhao Xuejun, Guo Guo, Wu Qinyi, Tao Ruyu, Wu Jianwei. Sequence Analysis,Cloning and Induced Expression of Chaperonin Gene in Housefly（Musca domesitca）[J]. Biotechnology Bulletin, 2014, 0(10): 151-155.

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