鸡法氏囊B淋巴细胞cDNA酵母表达文库的构建与鉴定  

Construction and identification of the yeast cDNA expression library of the B lymphocytes in bursa of Fabricius of chicken

作　　者：陈玉明[1] 高玉龙[1] 张礼洲[1] 王念[1] 高立[1] 任宪刚[1] 姜莉莉[1] 高宏雷[1] 秦立廷[1] 王永强[1] 祁小乐[1] 王笑梅[1] 

机构地区：[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,哈尔滨150001

出　　处：《黑龙江畜牧兽医》2014年第10期15-17,21,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：第43批教育部留学回国人员科研启动基金项目(无编号);国家重点实验室基本科研业务费项目(SKLVBP201303);现代农业产业技术体系建设专项基金项目(nycytx-42-G3-01)

摘　　要：为了研究传染性法氏囊病病毒(IBDV)与宿主B淋巴细胞作用机制,构建鸡法氏囊B淋巴细胞酵母表达文库,试验从3周龄SPF鸡中摘取法氏囊,分离制备B淋巴细胞,从中提取细胞总RNA,使用SMART和LD-PCR技术合成双链cDNA,并进一步与线性化载体pGADT7-Rec共同转化Y187酵母感受态细胞,通过同源重组构建酵母表达文库。结果表明:所构建cDNA文库的效价为9.5×107cfu/mL,重组率为100%,可以用于酵母双杂交试验。To study the mechanism of the interaction between infectious bursal disease virus （ IBDV ） and B lymphocyte in the host, and construct the yeast expression library of B lymphocyte in bursa of fabficius （BF）. The BF was obtained from the SPF chicken at 3 weeks of age, and then was isolated to prepare B lymphocyte. The total RNA of the B lymphocyte was extracted, and the double strains cDNA was synthesized using the techniques of SMART and LD - PCR. Furthermore, the cDNA and the linearized vector of pGADT7 - Rec were co - transformed into the Y187 yeast competent cells to construct the yeast expression library using homologous recombination. The results showed that the titer of con- structed cDNA library was 9.5 x 107 cfu/mL and its recombination rate was 100%. The results indicate that the yeast expression library can be used for the yeast two - hybrid experiment.

关 键 词：法氏囊 B淋巴细胞 酵母双杂交 CDNA文库 

分 类 号：S831.2[农业科学—畜牧学]