摘要
利用已测序的中国美利奴绵羊细菌人工染色体(BAC)文库MHC区段阳性克隆插入片段制备探针,筛选中国美利奴绵羊(新疆军垦型)混合组织cDNA文库,以期获得该MHC片段的基因组成与结构信息。用中国美利奴绵羊MHC ClassⅡb区域内的349I12 BAC克隆,BsaJⅠ酶切后制备α-32P放射性探针,以噬菌斑原位杂交筛选正常中国美利奴绵羊cDNA文库,并将分离所得的cDNA阳性克隆测序后进行基因结构分析。以349I12 BAC克隆制备探针经过两轮的噬菌斑原位杂交筛选,获得19个cDNA阳性克隆,经测序、比对等进一步分析确定获得17条不同序列,其中7条序列与免疫相关。绵羊20号染色体上的MHC区段包含表达序列,且多为断裂基因,对其基因结构的分析将有助于相应基因功能及调控方面的研究。
A sequenced clone in MHC region from Chinese Merino bacterial artificial chromosome（BAC） library was used to make probes for screening cDNA expression library of Chinese Merino sheep for some information about composition and structure of genes in the clone.The BAC clone 349I12 was located at the ClassⅡb domain of ovine major histocompatibility complex（MHC）.After digested by BsaJⅠ,the clone 349I12 was used to make probes with α-32P labeled.The probes were used to screen the Ovine cDNA library through phage in situ hybridization.The isolated positive cDNA clones were sequenced,then analyzed the composition and structure of genes from the BAC positive clone 349I12 by DNAstar.After screening twice,19 candidate positive cDNA clones were isolated using the probes from the clone 349I12.After further analysis through sequencing and GenBank inquiring,we finally got 17 pieces of different sequences,seven of them related to immunization.The MHC region located on No.20 chromosome contains MHC genes,immune related genes and others genes,lots of them being split genes.The analysis of gene structures would help in investigating genetic function and regulation of the genes.
出处
《西北农业学报》
CAS
CSCD
北大核心
2011年第8期6-11,共6页
Acta Agriculturae Boreali-occidentalis Sinica
基金
国家科技部国际合作重大项目(2006DFB33750)
国家"973"项目(2010CB530200)
