CLONING OF MELITTIN cDNA FROM HONEYBEE INTO THE HIGH EXPREHION PLASMID OF ESCHERICHIA COLI

• Published in: Insect science Vol. 4; no. 4; pp. 343 - 349
• Main Authors: Zhang, Qingwen, Chen, Zhongbin, Li, Jizhou, Cai, Qingnian, Zhang, Qin, Zhang, Fuling
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-12-1997
• Subjects: cDNA; clone; honeybee venom; mRNA
• ISSN: 1672-9609; 1744-7917
• DOI: 10.1111/j.1744-7917.1997.tb00108.x

Total mRNA from venom glands of newly emerged queen bees was reversely transcribed into cDNA and cloned into the EcoRI site of plasmid λgt11; cDNA library for bee venom was thus constructed. PCR technique was used to produce the melittin coding sequence from the cDNA library. A 87 bp product was produced and inserted into the EcoRI and PstI sites of the high level expression vector pBV220. Recombinant plasmid pBM95 was transformed into the competent cells of E.coli JM101. After screening transformants on LB medium with ampicilin, structure of the recombinant plasmid pBM95 from transformants was analyzed and melittin gene in pBM95 was sequenced. The cloned cDNA coding for honey bee melittin was obtained.