鹌鹑β- 防御素10基因的克隆与表达及其体内分布的检测  

Cloning and expression of quail avian beta-defensin 10 and the mRNA detection in quail tissues

作　　者：蔺利娟[1,2] 王瑞琴[1,2] 韩宗玺[2] 邵昱昊[2] 刘胜旺[2] 程宝晶[1] 孙黎[1] 李子瑞[1] 马得莹[1] 

机构地区：[1]东北农业大学动物营养研究所,黑龙江哈尔滨150030 [2]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,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2011年第7期552-556,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30972110);黑龙江省教育厅重点项目(1153LZ05)

摘　　要：为测定鹌鹑β-防御素10(AvBD10)体外抗菌活性及检测其在鹌鹑脏器组织的分布,本研究应用RT-PCR方法从鹌鹑肺脏组织中扩增到鹌鹑AvBD10,经序列同源性分析,鹌鹑AvBD10与鸡AvBD10的氨基酸序列同源性最高(83.8%)。将该基因亚克隆到表达载体pGEX-6p-1中进行原核表达,SDS-PAGE电泳表明,鹌鹑AvBD10重组蛋白分子量约为31ku。该重组蛋白经纯化后测定其体外抗菌活性,结果显示,鹌鹑AvBD10重组蛋白具有广谱的抗菌活性。采用定量RT-PCR法检测到AvBD10在鹌鹑脏器组织中广泛表达。To clone and express avian beta-defensin （AvBD10） gene in quail, the quail AvBD10 gene was amplified from lung of Chinese painted quail （Coturnix chinensis） by RT-PCR. Sequence analysis showed that the cDNA of quail AvBD10 consisted of 207 bp nucleotide, encoding a polypeptide of 68 amino acids. Homology analysis showed that quail AvBD10 shared 83.8% with chicken AvBD10. The cDNA of quail AvBD10 was sub-cloned into pGEX-6p-1 vector and expressed in E. coli BL21. SDS-PAGE analysis indicated that 31 ku of quail AvBD10 recombinant protein was highly expressed. The purified recombinant quail AvBD10 exhibit extensive antimicrobial activity against 10 different bacteria. Additionally, results of real-time RT-PCR detections showed that AvBD10 was widely expressed in various quail tissues.

关 键 词：鹌鹑 AvBD10 重组蛋白 抗菌活性 组织分布 

分 类 号：S852.72[农业科学—基础兽医学]