摘要
根据APD抗菌肽数据库报道的牛乳铁蛋白肽LfcinB氨基酸序列,合成编码LfcinB基因的2条互补的寡核苷酸链,退火后在5′端和3′端分别形成含有BamHI和XhoI位点粘性末端的双链DNA。真核表达载体pcDNA3.1(+)经BamHI和XhoI限制性内切酶双酶切处理后与合成的LfcinB基因进行连接,连接产物转化E.coliDH5α感受态细胞,提取质粒DNA进行测序。测序结果显示,牛乳铁蛋白肽LfcinB基因成功克隆到pcDNA3.1(+)真核表达载体中,为进一步表达和抗菌活性研究奠定物质基础。
Two complementary sense and antisense oligo-nucleotides, encoded bovine lactoferricin B gene （LfcinB）, were synthesized based on amino acids sequence of bovine lactoferricin B published in the antimicrobial peptide database （APD）. After the sense and antisense oligo-nucleotides annealed, a cohesive BamH Ⅰ site at 5＇terminus and a cohesive Xho Ⅰ site at 3＇terminus were formed. Synthesized LfcinB gene and eukaryotic expression vector pcDNA3.1 （＋）） digested with BamH Ⅰ and Xho I restriction endonuclease were ligated, and ligation products were transformed into E. coli DH5a. Plasmid extracted from transformed bacteria were sequenced by Sanger dideoxymethod. Sequencing result indicated the synthesized LfcinB gene was successfully cloned to multiple cloning site of vector pcDNA3.1 （＋）. This may lay a foundation for expression and antimicrobial activities in future.
出处
《中国畜牧兽医》
CAS
北大核心
2009年第5期95-98,共4页
China Animal Husbandry & Veterinary Medicine
关键词
牛乳铁蛋白
牛乳铁蛋白肽
真核表达载体
bovine lactoferrin
bovine lactoferricin B
eukaryotic expression vector
