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广东省自然科学基金(04020553)

作品数:2 被引量:6H指数:2
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家蚕拟抗微生物肽Gloverins基因(Bmglv)的原核表达及抗菌活性鉴定被引量:4
2008年
家蚕Bombyx mori基因组全测序于2004年完成后,由家蚕基因组注释发现了35条拟抗微生物肽基因序列。本研究选取其中的5条Gloverins同系物(isoforms)基因(BmglvB,BmglvA2,BmglvA3,BmglvA5和BmglvA6),克隆至pET-21d载体,转化Escherichia coli RosettaTM(DE3)宿主菌进行表达;克隆表达的5个家蚕Gloverins同系物基因大部分以可溶形式表达,经Ni-NTA亲和层析纯化和Sephadex G-10脱盐处理后,采用平板孔穴抑菌法测定其抗菌活性。结果表明:注释发现的5条家蚕拟抗微生物肽Gloverins同系物基因,具有与在其他昆虫中已鉴定报道的Gloverin相似的抗革兰氏阴性细菌的功能,实验确认了它们就是家蚕Gloverins抗微生物肽基因。
韩冬邓小娟杨婉莹叶明强曹方黄亚东温硕洋曹阳夏庆友
关键词:家蚕抗微生物肽原核表达抗菌活性
Secretive Expression of Insect Antifungal Peptide-Encoded Genes in Pichia pastoris and Activity Assay of the Products被引量:2
2007年
The antifungal peptides, drosomycin (Drs) and its isoform drosomycin-like C (Drs-lC) from Drosophila melanogaster and Thanatin from Podisus maculiventris, have potent activity with broad spectrum against filamentous fungi. Secretive expression of these genes in yeasts makes it possible to utilize the supernatants of yeast culture as protective reagents on fruit, vegetable, food and other agricultural products. So the study of effective secretion by yeast expression system is of great importance. Three genes, Drs, Drs-lC, and Thanatin, were cloned into pPICZαA and the recombinant vectors, pPICZαA-Drs, pPICZαA-Drs-lC, and pPICZαA-Thanatin were transformed into Pichia pastoris by the electric transfer method. The recombinant P. pastoris, which was screened by phenotype selection and PCR amplification, was induced to express antifungal peptide by methanol. The expressive products of the three recombinants showed antifungal activity against 5 out of 6 test fungi strains, and the products of Thanatin also had strong activity against the tested bacteria. The three antifungal peptide genes, Drs, Drs-lC, and Thanatin, were constructed into yeast P. pastoris. The expressed peptides were successfully secreted into the culture medium and exhibited potent activities against the test strains.
SANG Yan-xiaDENG Xiao-juanYANG Wan-yingWANG Wen-xianWEN Shuo-yangLIU Wen-quanHUANG Ya-dongCAO Yang
关键词:INSECT
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