摘要
绵羊主要组织相容性复合体(MHC)是与控制绵羊抗病性和易感性紧密连锁的基因簇。为了深入了解该类基因的组成与结构,利用中国美利奴绵羊细菌人工染色体(BAC)文库MHC区段克隆222G18,经BsaJⅠ酶切后制备α-32P放射性探针,通过噬菌斑原位杂交技术筛选中国美利奴绵羊cDNA文库,经过两轮杂交筛选,获得12个cDNA阳性克隆,经测序、比对等生物信息学分析确定获得7条与免疫相关的序列,其中3条具有完整的编码序列。利用SIM4软件将7条序列定位到BAC克隆上,结果显示绵羊MHC区段的表达序列多为断裂基因且跨度很大,可能是形成其基因多样性的重要原因之一。
The ovine major histocompatibility complex（MHC）harbors clusters of tightly linked genes involved in overall resistance/susceptibility of an animal to infectious pathogens.For a deeper understanding of composition and structure of such genes,a sequenced clone 222G18 in MHC region in Chinese Merino bacterial artificial chromosome（BAC）library was used to make probes with α-32P labeled after digested by BsaJ I.The probes were used to screen the cDNA library of Chinese Merino sheep through phage in situ hybridization.After screened twice,12 positive cDNA clones were isolated.After further analysis through sequencing and BLAST,we finally got 7 pieces of different sequences related to immunity,3 of them had complete coding sequences.The 7 pieces of sequences were located in BAC clone by SIM4 online and most of them were split genes and had long span structures.This may be one of the important reasons for the formation of the genetic diversity.The analysis of MHC gene structures would help in investigating function and application of the genes.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第12期127-131,共5页
Biotechnology Bulletin
基金
国家"973"项目子课题(2010CB530200)
