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| 牛乳腺炎无乳链球菌Pgk基因抗原优势区原核表达产物的抗原性分析 | |
| 引用本文： | 张海宝,布日额,吴金花,王学理,孙立杰,唐吉思,锡林高娃,刘燕,张忠祥.牛乳腺炎无乳链球菌Pgk基因抗原优势区原核表达产物的抗原性分析[J].中国兽医寄生虫病,2011,19(1):39-44. |
| 作者姓名： | 张海宝 布日额 吴金花 王学理 孙立杰 唐吉思 锡林高娃 刘燕 张忠祥 |
| 作者单位： | 1. 内蒙古民族大学生命科学学院,通辽,028043;内蒙古民族大学动物科技学院,通辽,028043 2. 内蒙古民族大学生命科学学院,通辽,028043 3. 内蒙古民族大学动物科技学院,通辽,028043 4. 内蒙古通辽市家畜繁育指导站,通辽,028000 |
| 基金项目： | 国家自然科学基金项目，内蒙古自治区高等学校科学技术研究项目 |
| 摘 要： | 为了探索牛乳腺炎无乳链球菌磷酸甘油酸激酶（Phosphoglycerate kinase,Pgk）基因编码蛋白的抗原性,根据GenBank公布的牛源无乳链球菌Pgk基因序列,设计并合成一对引物,通过PCR扩增出Pgk蛋白抗原优势区的编码序列,并对其进行克隆、测序、转化、诱导表达及抗原性鉴定。结果显示克隆的Pgk基因序列含有984 bp,编码328个氨基酸残基。临床分离株与GenBank上公布的B群无乳链球菌菌株Pgk基因（AE009948）核苷酸序列同源性为99.09%,氨基酸序列同源性为99.69%。将扩增的Pgk基因片段克隆至原核表达载体pET30a（＋）中,构建重组表达载体Pgk-pET30a（＋）,再将筛选的阳性重组质粒转化至BL21工程菌,经IPTG诱导获得部分可溶性表达Pgk重组蛋白。经Ni2＋亲和层析柱纯化得到纯度在90%以上的蛋白,经Western blot分析结果表明,重组蛋白具有良好的抗原性,为进一步探索其对奶牛的免疫原性研究奠定了良好的实验基础。 |
| 关 键 词： | 无乳链球菌 Pgk基因 原核表达 抗原性 |
ANTIGENICITY ANALYSIS OF PROKARYOTIC EXPRESSION PRODUCT OF PGK GENE ANTIGENIC DOMINANT REGION OF BOVINE MASTITIS STREPTOCOCCUS AGALACTIAE |
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| ZHANG Hai-bao,BU Ri-e,WANG Xue-li,WU Jin-hua,SUN Li-jie,TANG Ji-si,XI Lin-gao-wa,LIU Yan,ZHANG Zhong-xiang.ANTIGENICITY ANALYSIS OF PROKARYOTIC EXPRESSION PRODUCT OF PGK GENE ANTIGENIC DOMINANT REGION OF BOVINE MASTITIS STREPTOCOCCUS AGALACTIAE[J].Chinese Journal of Veterinary Parasitology,2011,19(1):39-44. | |
| Authors: | ZHANG Hai-bao BU Ri-e WANG Xue-li WU Jin-hua SUN Li-jie TANG Ji-si XI Lin-gao-wa LIU Yan ZHANG Zhong-xiang |
| Institution: | ZHANG Hai-bao1,2,BU Ri-e1,WANG Xue-li2,WU Jin-hua1,SUN Li-jie1,TANG Ji-si1,XI Lin-gao-wa1,LIU Yan1,ZHANG Zhong-xiang3 (1.College of Life Science,Inner Mongolia University for Nationalities,Tongliao 028043,China,2.College of Animal Science and Technology,Inner Mongolia University for Nationalities Tongliao 028043,3.Tongliao Guidance Station of Domestic Animal Reproduction,Tongliao 028000,China) |
| Abstract: | A pair of primers were designed and synthesized according to Pgk gene sequence of bovine mastitis streptococcus agalactiae published on Genbank.Antigenic dominant region of Pgk gene was amplified by PCR.The sequencing of PCR product showed that the cloned region of Pgk gene is 984bp in length,which encoding 328 amino acid residues.The nucleotide and amino acid identities of the partial Pgk gene between the clinical strain and Streptococcus agalactiae group B strain(AE009948) are 99.09% and 99.69%,respective... |
| Keywords: | Streptococcus agalactiae Pgk gene prokaryotic expression antigenicity |
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