鸡Gal-13成熟肽基因序列在毕赤酵母中的表达与抑菌活性分析
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- 英文论文题名：Expression of chickenβ-defensin-13mature peptide gene in Pichia pastoris and analysis of its antimicrobial activity
- 论文作者：许国洋 ; 付利芝 ; 杨金龙 ; 曹国文 ; 张素辉 ; 邱进杰 ; 潘康成
- 英文论文作者：XU Guo-yang ; FU Li-zhi ; YANG Jin-long ; CAO Guo-wen ; ZHANG Su-hui ; QIU Jin-jie ; PAN Kang-cheng ; China Sichuan Agricultural University ; Chongqing Academy of Animal Science
- 年：2016
- 作者机构：四川农业大学;重庆市畜牧科学院;
- 论文关键词：Gal-13 ; 成熟肽 ; 诱导表达 ; 抑菌活性
- 英文论文关键词：Gal-13 ; mature peptide ; inducible expression ; bacteriostatic activity
- 会议召开时间：2016-11-04
- 会议录名称：中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会论文集
- 语种：中文
- 分类号：Q78
- 学会代码：ZGXJ
- 会议名称：中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会
- 会议地点：中国山东青岛
- 主办单位：中国畜牧兽医学会动物微生态学分会
- 学会名称：中国畜牧兽医学会
- 页数：1
- 文件大小：71k
- 原文格式：O
- 会议级别：全国
摘要
扩增鸡β-防御素-13(Gal-13)成熟肽基因序列,构建酵母重组表达载体pPICZαA-Gal-13,通过Sac I限制性内切酶线性化处理后,电击转化毕赤酵母X-33,并对利用甲醇诱导后的重组菌株表达产物进行表达规律分析和抑菌活性研究。结果表明,扩增到的Gal-13成熟肽基因序列为108bp,共编码36个氨基酸;筛选到的重组菌株均为Mut+表型,其诱导表达上清经Tricine-SDS-PAGE凝胶电泳后,发现其在约5900的位置出现目的条带,抑菌试验发现其对沙门菌(CVCC534)和金黄色葡萄球菌(ATCC25923)具有抑菌活性。
In order to analyze the expression principle and the bacteriostatic activity of chicken β-defensin-13(Gal-13),the mature peptide gene sequence of Gal-13 was augmented and inserted into pPICZaA to build the recombinant plasmid pPICZαA-Ga1-13,and the correct recombinant plasmid was linearized and transformed into Pichia Pastoris strain X-33 by electroporation.The result showed that the mature peptide gene of Gal-13 consists of 312 bp,encoding 36 amino acids.The recombination strain,which was induced by methyl alcohol,was Mut~+ phenotype,and the target protein secreted into the microbial supernatant showed at a level of 5.9 ku according to the Tricine-SDS-PAGE.The antimicrobial bioactivity tests showed that Gal-13 had antimicrobial bioactivity against Salmonella gallinarum(CVCC534) and Staphylococcus aureus(ATCC25923).
In order to analyze the expression principle and the bacteriostatic activity of chicken β-defensin-13(Gal-13),the mature peptide gene sequence of Gal-13 was augmented and inserted into pPICZaA to build the recombinant plasmid pPICZαA-Ga1-13,and the correct recombinant plasmid was linearized and transformed into Pichia Pastoris strain X-33 by electroporation.The result showed that the mature peptide gene of Gal-13 consists of 312 bp,encoding 36 amino acids.The recombination strain,which was induced by methyl alcohol,was Mut~+ phenotype,and the target protein secreted into the microbial supernatant showed at a level of 5.9 ku according to the Tricine-SDS-PAGE.The antimicrobial bioactivity tests showed that Gal-13 had antimicrobial bioactivity against Salmonella gallinarum(CVCC534) and Staphylococcus aureus(ATCC25923).
引文
[1]王海英,祁克宗,彭开松.禽防御素代替抗生素最为饲料添加剂的研究现状[J].兽药与饲料添加剂,2008,13(5):27-28.
[2]李斌,云才,肖永仁。抗菌肤及其在畜牧生产中的应用[J].吉林畜牧兽医,2005(10):15-17.
[3]Harwig S S,Swiderek K M,Kokryakov V N,et al.Gallinacins:cysteine-rich antimicrobial peptides of chicken leukocytes[J].FEBS Lett,1994,342(3):281-285.
[4]Evans E w,Beach G G,wunderlich J,et al.Isolation of antimicrobial peptides from avian heterophils[J].J Leukoc Bi01,1994,56(5):661-665.
[5]王新卫,康相涛.禽类防御素研究进展[J].动物医学进展,2006,27(12):35-37.
[6]Ezzine A,M'hirsi El Adab S,Bouhaouala-Zahar B,et al.Efficient expression of the anti-Aahl'scorpion toxin nanobody under a new functional form in a Pichia pastoris system[J].Biotechnol Appl Biochem,2012,59(1):15-21.
[2]李斌,云才,肖永仁。抗菌肤及其在畜牧生产中的应用[J].吉林畜牧兽医,2005(10):15-17.
[3]Harwig S S,Swiderek K M,Kokryakov V N,et al.Gallinacins:cysteine-rich antimicrobial peptides of chicken leukocytes[J].FEBS Lett,1994,342(3):281-285.
[4]Evans E w,Beach G G,wunderlich J,et al.Isolation of antimicrobial peptides from avian heterophils[J].J Leukoc Bi01,1994,56(5):661-665.
[5]王新卫,康相涛.禽类防御素研究进展[J].动物医学进展,2006,27(12):35-37.
[6]Ezzine A,M'hirsi El Adab S,Bouhaouala-Zahar B,et al.Efficient expression of the anti-Aahl'scorpion toxin nanobody under a new functional form in a Pichia pastoris system[J].Biotechnol Appl Biochem,2012,59(1):15-21.