| Gallinacin-2在毕赤酵母中的分泌表达 |
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| 引用本文： | 王强,彭开松,严兵,涂健,屠利民,祁克宗.Gallinacin-2在毕赤酵母中的分泌表达[J].生物学杂志,2011,28(2):43-46. |
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| 作者姓名： | 王强 彭开松 严兵 涂健 屠利民 祁克宗 |
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| 作者单位： | 安徽农业大学动物科技学院动物分子病理室,合肥,230036 |
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| 基金项目： | 国家高技术研究发展计划(863)项目，国家自然科学基金 |
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| 摘 要： | 利用RT-PCR技术,从鸡肺脏总RNA中克隆了Gal-2 cDNA全序列,以克隆载体为模板PCR扩增Gal-2成熟肽cDNA,将成熟肽cDNA插入到分泌型表达载体pPIC9k中,构建pPIC9k-Gal-2成熟肽表达载体;通过电转化方法将表达质粒导入到毕赤酵母GS115体内,阳性克隆菌经甲醇诱导表达,SDS-PAGE蛋白电泳检测在3.9ku附近出现预期大小的条带,灰度扫描显示蛋白表达量占总分泌蛋白量的63.7%。抑菌试验结果表明,表达的防御素对大肠埃希氏菌、乙型副伤寒沙门氏菌、枯草芽孢杆菌和金黄色葡萄球菌有一定的抑菌活性。
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| 关 键 词： | β-防御素 Gal-2 pPIC9k 毕赤酵母 |
Secreted expression of Gallinacin-2 in Pichia pastoris |
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| WANG Qiang,PENG Kai-song,YAN Bing,TU Jian,TU Li-ming,Qi Ke-zong.Secreted expression of Gallinacin-2 in Pichia pastoris[J].Journal of Biology,2011,28(2):43-46. |
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| Authors: | WANG Qiang PENG Kai-song YAN Bing TU Jian TU Li-ming Qi Ke-zong |
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| Institution: | (Animal Molecular Pathology Room,College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China) |
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| Abstract: | A full-length Gal-2 cDNA was amplified by RT-PCR from total RNA of chickren lung,the mature cDNA of Gal-2 was amplified from the clone plasmid,and then cloned into the secreted expression plasmid pPIC9k.The recombinant vector pPIC9k-Gal-2 was transformed by electroporation into Pichia pastoris GS115.The positive clones were cultured and induced to express target protein by methanol.The expression products were approximately 3.9ku in molecular weight,and they were examined by SDS-PAGE.Densitometric scanning analysis demonstrated that the expressed protein exaccounted for about 63.7% of the total secreted bacterial protein.The expression products had antimicrobial activity against E coli,S.paratyphi B,B subtilis and S aureus. |
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| Keywords: | Gal-2 pPIC9k |
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