Molecular Cloning and Prokaryotic Expression of Prothoracicotropic Hormone cDNA Sequence from Heliothis viriplaca

Abstract

Abstract: Prothoracicotropic hormone（PTTH）is a group of important insect hormones. They have vital effect on regulation of growth, ecdysis, metamorphosis and diapause of insects. Total RNA was isolated from the fifth instar larvae of Heliothis viriplaca. The cDNA sequence was cloned by RT-PCR and rapid amplification of cDNA ends（RACE）. The cDNA sequence was 823 base pairs in length and contained an open reading frame of 681 base pairs, encoding for a polypeptide of 226 amino acid residues, with a predicted molecular weight of 26.3 kD and pI 8.51. Sequence analysis showed that the predicted amino acid shared extensive similarities with those from other insects, especially the lepidopteran noctuid insects. The cDNA sequence has been deposited in GenBank with accession No. JF731347. The cDNA sequence was successfully expressed by the recombinant prokaryotic expression vector pET21b in E. coli. The recombinant protein with His-tag was purified by Ni-NTA affinity chromatography.

Key words: Heliothis viriplaca, Prothoracicotropic hormone, cDNA cloning, Prokaryotic expression

Zhou Xia, Guo Bozhi, Gao Yanling, Zhao Kuijun, Fan Dong. Molecular Cloning and Prokaryotic Expression of Prothoracicotropic Hormone cDNA Sequence from Heliothis viriplaca[J]. Biotechnology Bulletin, 2014, 0(10): 144-150.

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