湖北农业科学
湖北農業科學
호북농업과학
2014年
7期
1684-1689
,共6页
李一星%吴梅%李友国
李一星%吳梅%李友國
리일성%오매%리우국
紫云英(Astragalus sinicus)%共生固氮%抑制消减杂交%下调表达基因
紫雲英(Astragalus sinicus)%共生固氮%抑製消減雜交%下調錶達基因
자운영(Astragalus sinicus)%공생고담%억제소감잡교%하조표체기인
Astragalus sinicus%symbiotic nitrogen fixation%suppressive subtractive hybridization%down-regulated gene
对前期利用抑制消减杂交技术（Suppressive subtractive hybridization， SSH)构建的紫云英（A stra-galus sinicus)根部组织的共生固氮差异表达cDNA文库进行了全文库测序和初步分析，并利用半定量RT-PCR方法验证了其中5个基因片段在根瘤中的下调表达。结果表明，从180个克隆中共获得94个有效序列，文库插入片段长度为200~1300 bp。 BLAST同源比对分析结果表明，有90个序列可以找到同源片段，有4个可能为新基因，按功能分为10个类群。
對前期利用抑製消減雜交技術（Suppressive subtractive hybridization， SSH)構建的紫雲英（A stra-galus sinicus)根部組織的共生固氮差異錶達cDNA文庫進行瞭全文庫測序和初步分析，併利用半定量RT-PCR方法驗證瞭其中5箇基因片段在根瘤中的下調錶達。結果錶明，從180箇剋隆中共穫得94箇有效序列，文庫插入片段長度為200~1300 bp。 BLAST同源比對分析結果錶明，有90箇序列可以找到同源片段，有4箇可能為新基因，按功能分為10箇類群。
대전기이용억제소감잡교기술（Suppressive subtractive hybridization， SSH)구건적자운영（A stra-galus sinicus)근부조직적공생고담차이표체cDNA문고진행료전문고측서화초보분석，병이용반정량RT-PCR방법험증료기중5개기인편단재근류중적하조표체。결과표명，종180개극륭중공획득94개유효서렬，문고삽입편단장도위200~1300 bp。 BLAST동원비대분석결과표명，유90개서렬가이조도동원편단，유4개가능위신기인，안공능분위10개류군。
In previous work, a cDNA library of Astragalus sinicus genes related with symbiotic nitrogen fixation was con-structed with suppressive subtractive hybridization(SSH﹚. The library containing down-regulated genes was sequenced and an-alyzed. Among the sequenced genes, five fragments were sckeened and its down-regulated expressions were confirmed by se-mi-quantitative RT-PCR. The results showed that 94 genes were obtained from 180 clones and the average length of inserted fragments was 200~1 300 bp. Nucleotide BLAST homological analysis showed that 90 genes had similarities to known genes and 4 genes were putatively novel genes. All the 94 genes were divided into 10 functional categories.
