摘要
本研究所构建的BAC文库覆盖了8倍新疆细毛羊的基因组，平均插入片段的大小为133kb．同时文库92．5％的克隆插入片段大于100kb，而且有部分克隆甚至大于300kb，假定绵羊的基因组含有3×10^6kb，根据文库的平均插入片段大小为133kb，从文库筛选到目的片段的概率为98．208％。为了验证文库有较好的覆盖率．构建了2倍基因组文库PCR筛选系统，并对位于新疆细毛羊20号染色体MHC基因邻近区段的DMB_EX2、MCMA36、CP73和BM12584个分子标记进行了筛选．得到的平均阳性克隆数为1．5个，从筛选结果来看，这与文库插入片段估计的8倍基因组覆盖率相当接近并且没有偏向，这使得本文库成为研究绵羊的功能基因、位置克隆和完善基因组物理图谱的极为有用的资源。
In this study,we used our special regional fostering breed Xinjiang fine-fleece sheep＇s genome to construct its genome library. The average size of insert fragments of the library was 133 kb. At the same time 92.5% clones＇ insert fragments of the library were larger than 100 kb and some larger than 300 kb. If we supposed sheep＇s genome contenting 3×10^6 kilo-base, on the basis of the average insert fragment was 133 kb the library covered 8 times genome of Xinjiang fine-fleece sheep. The probability of the tagged fragment being screened from the library was 98.208%. To prove the BAC library had been the better rate of coverage,four molecular markers： DMB_EX2,MCMA36,CP73 and BM1258 located to MHC gene of chromosome 20 near region in xinjiang fine-fleece sheep had been screened positive clones from the constructed 2 times genome library PCR screening system and the average positive clones was 1.5. The screening result showed that the constructed genome library was fairly closed to the 8 times genome coverage and had no erroneous tendency, which made the library being of the utmost useful resource for studying functional gene, position cloning and improving the genome physical map of sheep.
出处
《分子细胞生物学报》
CSCD
北大核心
2006年第6期489-494,共6页
Journal of Molecular Cell Biology
基金
国家重大基础研究前期研究专项基金(2002CCC2100)。~~
