抗菌肽CAP18在大肠杆菌中表达、纯化及活性的初步鉴定

被引：2

作者：

徐灵龙 ^{[1
,2
]}

石星明 ^{[1
]}

王云峰 ^{[1
]}

史春林 ^{[1
]}

王玫 ^{[1
]}

孙妍 ^{[1
]}

童光志 ^{[3
]}

机构：

[1] 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽传染病研究室

[2] 东北农业大学生命科学学院

[3] 中国农业科学院上海兽医研究所

来源：

中国预防兽医学报 | 2008年 / 11期

关键词：

兔抗菌肽CAP18; 融合基因; 重组表达; 抑菌活性;

D O I：

暂无

中图分类号：

Q78 [基因工程（遗传工程）];

学科分类号：

071007 [遗传学];

摘要：

根据抗菌肽CAP18活性片段的氨基酸序列,采用SOE技术,设计合成了CAP18的编码基因,将其克隆至pMD18-T载体,进行序列测定后亚克隆至pET-32a(+)表达载体中,在E.coli BL21(DE3)中表达融合蛋白CAP18。经Ni2+亲和层析后回收CAP18融合蛋白,纯度达到了85%以上。对纯化的融合蛋白进行酶切,用液体生长抑制方法检测切割后样品的活性,结果表明重组抗菌肽CAP18对大肠杆菌具有抑制活性。

引用

页码：861 / 865

页数：5

共 8 条

[1]

Preparation of recombinant thioredoxin fused N-terminal proCNP: Analysis of enterokinase cleavage products reveals new enterokinase cleavage sites [J].

Liew, OW ;

Chong, JPC ;

Yandle, TG ;

Brennan, SO .

PROTEIN EXPRESSION AND PURIFICATION, 2005, 41 (02) :332-340

[2]

Efficient site-specific processing of fusion proteins by tobacco vein mottling virus protease in vivo and in vitro [J].

Nallamsetty, S ;

Kapust, RB ;

Tözsér, J ;

Cherry, S ;

Tropea, JE ;

Copeland, TD ;

Waugh, DS .

PROTEIN EXPRESSION AND PURIFICATION, 2004, 38 (01) :108-115

[3]

Expression, purification, and characterization of human enteropeptidase catalytic subunit in Escherichia coli [J].

Gasparian, ME ;

Ostapchenko, VG ;

Schulga, AA ;

Dolgikh, DA ;

Kirpichnikov, MP .

PROTEIN EXPRESSION AND PURIFICATION, 2003, 31 (01) :133-139

[4]

Recombinant enterokinase light chain with affinity tag:: Expression from Saccharomyces cerevisiae and its utilities in fusion protein technology [J].

Choi, SI ;

Song, HW ;

Moon, JW ;

Seong, BL .

BIOTECHNOLOGY AND BIOENGINEERING, 2001, 75 (06) :718-724

[5]

Influence of the amino acid residue downstream of (Asp)4Lys on enterokinase cleavage of a fusion protein [J].

Hosfield, T ;

Lu, Q .

ANALYTICAL BIOCHEMISTRY, 1999, 269 (01) :10-16

[6]

Codon optimization of the gene encoding a domain from human type 1 neurofibromin protein results in a threefold improvement in expression level in Escherichia coli [J].

Hale, RS ;

Thompson, G .

PROTEIN EXPRESSION AND PURIFICATION, 1998, 12 (02) :185-188

[7]

Expression and Purification of a Recombinant Tobacco Etch Virus NIa Proteinase: Biochemical Analyses of the Full-Length and a Naturally Occurring Truncated Proteinase Form.[J].T.Dawn Parks;Eric D. Howard;Thomas J. Wolpert;Daniel J. Arp;William G. Dougherty.Virology.1995, 1

[8]

Defensins: Antimicrobial and Cytotoxic Peptides of Mammalian Cells.[J].R I Lehrer;A K Lichtenstein;T Ganz.Annual Review of Immunology.1993,

← 1 →