GLGI技术鉴定奶山羊乳腺中的κ-酪蛋白基因  

Identification of κ-casein gene by GLGI technique in long-sAGE tags of dairy goat mammary gland

作　　者：孙瑞秋[1] 赵婧[1] 闫宏博[1] 李庆章[1] 高学军[1] 

机构地区：[1]东北农业大学乳品科学教育部重点实验室,哈尔滨150030

出　　处：《中国乳品工业》2010年第12期4-7,共4页China Dairy Industry

基　　金：黑龙江省国际合作项目;东北农业大学创新团队项目(CXT005-1-1)

摘　　要：采用GLGI技术扩增本实验室构建的Long-SAGE标签库中的κ-酪蛋白基因。使用SMARTTM RACE cDNA Ampliication Kit通过RT-PCR把奶山羊乳腺组织中的mRNA逆转录成cDNA,分别用3’RACE和5’RACE扩增κ-酪蛋白基因的3’端和5’端。扩增产物经TA克隆后转化大肠杆菌,随机挑取阳性克隆后进行测序分析。通过序列比对、拼接,获得全长的κ-酪蛋白基因。应用GLGI技术成功获得了完整的奶山羊乳腺CSN3序列,为CSN3结构分析和功能研究奠定了重要基础。An optimized procedure of GLGI was adopted to amplify κ-casein gene sequence in Long-SAGE tags of dairy goat mammary gland.SMARTTM RACE cDNA Ampliication Kit was used to mRNA of dairy goat mammary gland tissue reverse transcribe to cDNA by RT-PCR.3′ and 5＇ cDNA fragments of κ-casein gene were extended by 3＇RACE and 5＇RACE respectively.Products of PCR were T/A-cloned and transformed to E.coli.Positive colonies were sequenced.Total length κ-casein gene was obtained by sequence contrasting and splicing.Complete CSN3 sequence of dairy goat mammary gland were successfully identified by applying this high-throughput procedure,which established a basis for further structural and functional analysis of CSN3.

关 键 词：GLGI RACE Κ-酪蛋白基因 

分 类 号：Q93-33[生物学—微生物学]