In order to study the expression of function gene and its effect on metabolic control and other physiological function in liver, 438 expressed sequence tags (ESTs) were determined, which were from a cD-NA library of p...In order to study the expression of function gene and its effect on metabolic control and other physiological function in liver, 438 expressed sequence tags (ESTs) were determined, which were from a cD-NA library of porcine liver tissue. The results showed that the nucleotide sequences of 186 ESTs have already presented in GenBank database, and 37 ESTs could be found the homology with human and other species, while the others were not identified. 45 full length insertion of the clones randomly isolated from cDNA library were also completely sequenced with different size, and the results showed that 19 of them were function-known genes, 11 had no open reading frame ( ORF )at all and 15 had ORF but the function were not elucidated yet.展开更多
目的构建亚洲带绦虫成虫全长cDNA质粒文库,为获取亚洲绦虫成虫的基因信息,建立基因表达谱,并为筛选疫苗基因和诊断抗原基因奠定基础。方法提取亚洲带绦虫成虫mRNA,构建pBluescript II SK全长cDNA质粒文库,测定扩增文库的滴度;用载体克...目的构建亚洲带绦虫成虫全长cDNA质粒文库,为获取亚洲绦虫成虫的基因信息,建立基因表达谱,并为筛选疫苗基因和诊断抗原基因奠定基础。方法提取亚洲带绦虫成虫mRNA,构建pBluescript II SK全长cDNA质粒文库,测定扩增文库的滴度;用载体克隆位点两端的引物进行PCR扩增,以检测所构建文库的质量。随机挑选质粒文库转化的阳性重组克隆,进行较大规模的5′端测序,归并unigene。结果测定文库的滴度为1011pfu/μl,插入片段的大小主要在1000bp以上。共测1495个克隆,获得有效EST序列1126个,归并为643条unigene。结论已成功获得一高质量的亚洲带绦虫成虫全长cDNA表达文库,并获得了较丰富的成虫表达基因数据。展开更多
以核盘菌子囊盘为材料,应用SMART(Switch Mechanism at 5′end of RNA Transcript)技术,构建了高质量的核盘菌cDNA文库.涂平板测定和酶切反应鉴定表明,原始文库的滴度为4.11×105,重组率达97.3%.通过对文库克隆的序列测定和初步生...以核盘菌子囊盘为材料,应用SMART(Switch Mechanism at 5′end of RNA Transcript)技术,构建了高质量的核盘菌cDNA文库.涂平板测定和酶切反应鉴定表明,原始文库的滴度为4.11×105,重组率达97.3%.通过对文库克隆的序列测定和初步生物信息学分析,共获得1800条核盘菌有效表达序列标签(Expressed Sequence Tag,EST).同源比对分析表明,844条独立基因代表了242个已知基因,其余602个EST为未知序列.这242个已知基因大致可以分为3类:78个可归属于生物学过程,97个可归属于分子功能,67个可归属于细胞组分.其中,在97个分子功能基因中筛选到1个与MADS-box具有较高同源性的基因(SsMADS),推测可能与核盘菌的有性生殖过程有关.展开更多
基金supported by the Chinese National Key Project of Basic Research(G20000161)
文摘In order to study the expression of function gene and its effect on metabolic control and other physiological function in liver, 438 expressed sequence tags (ESTs) were determined, which were from a cD-NA library of porcine liver tissue. The results showed that the nucleotide sequences of 186 ESTs have already presented in GenBank database, and 37 ESTs could be found the homology with human and other species, while the others were not identified. 45 full length insertion of the clones randomly isolated from cDNA library were also completely sequenced with different size, and the results showed that 19 of them were function-known genes, 11 had no open reading frame ( ORF )at all and 15 had ORF but the function were not elucidated yet.
文摘目的构建亚洲带绦虫成虫全长cDNA质粒文库,为获取亚洲绦虫成虫的基因信息,建立基因表达谱,并为筛选疫苗基因和诊断抗原基因奠定基础。方法提取亚洲带绦虫成虫mRNA,构建pBluescript II SK全长cDNA质粒文库,测定扩增文库的滴度;用载体克隆位点两端的引物进行PCR扩增,以检测所构建文库的质量。随机挑选质粒文库转化的阳性重组克隆,进行较大规模的5′端测序,归并unigene。结果测定文库的滴度为1011pfu/μl,插入片段的大小主要在1000bp以上。共测1495个克隆,获得有效EST序列1126个,归并为643条unigene。结论已成功获得一高质量的亚洲带绦虫成虫全长cDNA表达文库,并获得了较丰富的成虫表达基因数据。
文摘以核盘菌子囊盘为材料,应用SMART(Switch Mechanism at 5′end of RNA Transcript)技术,构建了高质量的核盘菌cDNA文库.涂平板测定和酶切反应鉴定表明,原始文库的滴度为4.11×105,重组率达97.3%.通过对文库克隆的序列测定和初步生物信息学分析,共获得1800条核盘菌有效表达序列标签(Expressed Sequence Tag,EST).同源比对分析表明,844条独立基因代表了242个已知基因,其余602个EST为未知序列.这242个已知基因大致可以分为3类:78个可归属于生物学过程,97个可归属于分子功能,67个可归属于细胞组分.其中,在97个分子功能基因中筛选到1个与MADS-box具有较高同源性的基因(SsMADS),推测可能与核盘菌的有性生殖过程有关.
