摘要
为构建猪PBD-1基因乳腺特异性表达载体,采用PCR方法从质粒pcDNA3.1-BLG-HNP-1中扩增出山羊乳球蛋白(BLG)基因,插入到真核表达载体pIRES2-EGFP-PBD-1构建成乳腺特异性表达载体pIRES2-EGFP-BLG-PBD-1。经PCR鉴定、限制性内切酶酶切分析和克隆片段序列测定、比较,鉴定。结果成功构建了乳腺特异性表达载体pIRES2-EGFP-BLG-PBD-1。乳腺特异性表达载体pIRES2-EGFP-BLG-PBD-1的成功构建,为进一步研究PBD-1蛋白的抗菌活性、抗菌机理及将进一步该载体应用于动物乳腺生物反应器的研究奠定基础。
It was to construct a mammary gland special expression vector for the porcine beta-defensin 1. BLG gene was amplified from the vector of pcDNA3.1-BLG-HNP-1 and inserted into a eukaryotic expression plasmid pIRES2-EGFP-PBD-1. The recombinant colonies were identified by the methods of restriction enzyme digestion, PCR and sequencing. Results showed that the mammary gland special expression vector pIRES2-EGFP-BLG-PBD-1 was successfully constructed. The establishment of mammary gland special expres- sion vector pIRES2-EGFP-BLG-PBD-1 would lays a foundation in research on antimicrobial activities and its mechanism of the de- fensins, and also would benefit further research on the establishment of animal mammary bioreactor.
出处
《生物技术通报》
CAS
CSCD
北大核心
2009年第5期127-129,134,共4页
Biotechnology Bulletin
基金
国家“863”课题(2007AA100505)
武汉市科技攻关计划(200820422192)
