CAJ | 학술논문

利用SMART cDNA文库构建试剂盒构建了文蛤(Meretrix meretrix)肠、外套膜和肝胰脏组织的cDNA文库.经测定原始文库滴度分别为2.10×10~6、1.70×10~6和1.60×10~6,重组率高于95%,肠组织文库插入片段长度均大于1 000bp,外套膜和肝胰脏组织文库的插入片段长度大于1 kb的占87.5%,文库质量符合标准要求.随机选取文库的3 168个克隆进行5'端测序,获得高质量表达序列标签(Expressed Sequence Tags,ESTs)3029个(肠:1 005,外套膜:1 019,肝胰脏:1 005),测序成功率95.58%.经质量控制和拼接得到1 796个单基因簇(Unigene),其中306个叠联群(Contigs),1 490个单一序列(Singletons).通过Blastx搜索比对、查询和注释分析,共得到已知基因696个(肠:216,外套膜:235个;肝胰脏:245个),占总数38.75%.在1 796个单基因簇(Unigene)发现微卫星序列55条,这些存在微卫星位点的序列占整个ESTs数据库的3.1%.
이용SMART cDNA문고구건시제합구건료문합(Meretrix meretrix)장、외투막화간이장조직적cDNA문고.경측정원시문고적도분별위2.10×10~6、1.70×10~6화1.60×10~6,중조솔고우95%,장조직문고삽입편단장도균대우1 000bp,외투막화간이장조직문고적삽입편단장도대우1 kb적점87.5%,문고질량부합표준요구.수궤선취문고적3 168개극륭진행5'단측서,획득고질량표체서렬표첨(Expressed Sequence Tags,ESTs)3029개(장:1 005,외투막:1 019,간이장:1 005),측서성공솔95.58%.경질량공제화병접득도1 796개단기인족(Unigene),기중306개첩련군(Contigs),1 490개단일서렬(Singletons).통과Blastx수색비대、사순화주석분석,공득도이지기인696개(장:216,외투막:235개;간이장:245개),점총수38.75%.재1 796개단기인족(Unigene)발현미위성서렬55조,저사존재미위성위점적서렬점정개ESTs수거고적3.1%.
The cDNA libraries of intestine, mantle and hepatopancreas of hard clam (Meretrix meretrix)were constructed with SMART cDNA Library Construction Kit. The libraries have a high titer of 2.10×10~6, 1.70×10~6 and 1.60×10~6, respectively. The recombined efficiency of each cDNA library exceeded 95%. The all insert sizes of intestine library were over 1 kb, and the insert sizes over 1 kb in the mantle and hepatopancreas libraries were over 87.5%. Atotal of 3 168 clones of the libraries were randomly picked and sequenced from the 5' end of the cDNAs using a M13 universal primer, and 3 029 raw sequences of the ESTs were processed and then assembled into 1 796 unigene involving 306 contigs and 1 490 singletons. Blastx analysis showed that 696 unigenes (216 for intestine, 235 for mantle and 245 for hepatopancreas) (38.75%) had significant homology (Evalue ≤10~(-5)) to genes with known or putative functions in GenBank.The sequences containing microsatellite DNA (SSR) accounted for 3.1% of all ESTs indicating that SSR sequences were rich in the ESTs of hard clam.