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Title of Journal: Mol Biol Rep

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Abbravation: Molecular Biology Reports

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Springer Netherlands

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DOI

10.1016/0003-6870(84)90083-8

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ISSN

1573-4978

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Molecular analysis and recombinant expression of b

Authors: Jianming Wu Changfa Wang Hongbin He Guixue Hu Hongjun Yang Yundong Gao Jifeng Zhong
Publish Date: 2010/03/28
Volume: 38, Issue: 1, Pages: 429-436
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Abstract

To analyze molecular characteristics and antimicrobial activity of bovine neutrophil βdefensin 12 BNBD12 ProBNBD12 gene was amplified from Chinese Holstein cow using reverse transcription polymerase chain reaction Based on the sequence of mature BNBD12 and codon preference in E coli a modified mature BNBD12 cDNA was synthesized cloned into pET32a + vector and expressed in E coli BL21 as a 26 kD fusion protein after isopropyl βd1thiogalactopyranoside induction The expressed mature BNBD12 accounted for 214 of total protein 005 mg/ml purified BNBD12 had antimicrobial activity against both E coli and S aureus in vitro assayed by agar diffusion method Electron microscopy found that BNBD12 treatment of E coli and S aureus could induce cell content leakage Taking together BNBD12 protein was successfully expressed in E coli and it has antimicrobial activity against both Grampositive and negative bacteria and potentials in control of mastitis clinicallyThis work is supported by “863” project of the Ministry of Science and Technology P R China No 2007AA10Z169 and 2006AA10Z1D9 Program of National Cow Industrial Technology System No nycytx0107 AntiTB Transgenic Project 2008ZX08007004 and Young Scientist Fund in Shandong Province 2006BS06010

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