海带雄性配子体差异表达基因片段的克隆及筛选-水产学报.PDF

29 5 Vol. 29, No. 5 2005 10 JOURNAL OF FISHERIES OF CHINA Oct. , 2005 : 1000- 0615( 2005) 05- 0666- 04 史西志, 毕燕会, 周志刚 ( , , 200090) : ( uppre ion ubtractive hybridization, SSH) cDNA , cDNA pGEM- T ( Escherichia coli) JM109, cDNA 750 100 PCR , 200~ 1000 bp Southern , 10 , 7 GenBank , , 3 Fcp Lhcf 6 88% 98% : ; ; ; cDNA ; : S917 : A Cloning and screening of differentially expressed genes from the male gametophytes of Laminaria j ap onica Aresch SHI Xi-zhi, BI Yan-hui, ZHOU Zh-i gang (Key Laboratory of A quatic Genetic Resources andA quacultural Ecology Certfi icated by the Ministry of Agriculture, College of Aqua-lfie cience and Technology, hanghai Fisheries University, hanghai 200090, China) Abstract: Differentially expre ed cDNA ubtracted female from male gametophyte of Laminaria j aponica were obtained by uppre ion ubtractive hybridization ( SSH) . The differentially expre ed cDNA from the male gametophyte were ligated into the pGEM-T vector and the latter wa con equentially tran formed into Escherichia coli JM 109 competent cell . The ubtracted cDNA librarie of male gametophyte were con tructed. One hundred randomly elected clone were amplified by PCR u ing T7 and SP6 primer for primary creening . The length of in erted fragment a ayed by PCR amplification wa mo tly from 200 to 1000 ba e pair . Ten clone obtained after do-t blot Southern hybridization were equenced and analyzed for homology compari on u ing BLA ST. Seven of the ten clone howed no equence homology in the GenBank databa e, ugge ting that they or at lea t ome of them with