CAJ | 학술논문

以苦瓜(Momordica charantia L.)1/4黄期果实为材料,采用DSN (duplex-specific nuclease)均一化与SMART (switching mechanism at 5' end of RNA transcript)技术相结合,构建成熟软化期苦瓜果实均一化全长cDNA文库.经检测,该文库的转化效率为3600000个菌落/μL连接产物,其平均插入片段大小为1.4 kb,重组率为98.0%,表明文库质量较好.随机挑取768个克隆进行EST测序,获得有607个高质量的EST,其中17个contig和562个singleton.冗余率为7.41%,文库的全长率67%.对测序结果进行注释和归类分析表明,已知功能基因大部分与催化活性、蛋白结合、转录调节活性、转运活性和酶调控活性有关分析,结果表明文库的可靠性好.发现有若干个与胞内蛋白质降解、细胞壁代谢、乙烯合成、信号传导和抗逆等可能与苦瓜果实成熟软化相关的新基因.
이고과(Momordica charantia L.)1/4황기과실위재료,채용DSN (duplex-specific nuclease)균일화여SMART (switching mechanism at 5' end of RNA transcript)기술상결합,구건성숙연화기고과과실균일화전장cDNA문고.경검측,해문고적전화효솔위3600000개균락/μL련접산물,기평균삽입편단대소위1.4 kb,중조솔위98.0%,표명문고질량교호.수궤도취768개극륭진행EST측서,획득유607개고질량적EST,기중17개contig화562개singleton.용여솔위7.41%,문고적전장솔67%.대측서결과진행주석화귀류분석표명,이지공능기인대부분여최화활성、단백결합、전록조절활성、전운활성화매조공활성유관분석,결과표명문고적가고성호.발현유약간개여포내단백질강해、세포벽대사、을희합성、신호전도화항역등가능여고과과실성숙연화상관적신기인.
@@@@A normalized full-length cDNA library was constructed from the bitter gourd fruits during in its ripening and softening stage, by DSN normalization method combined with SMART technique. The Conversion efficiency of original cDNA library was about 3 600 000 colonies/μL. The average size of inserted fragments was more 1.4 kb. Random picking clones showed that the recombination rate was about 98%. The results suggested that the quality of the library be better. Random selected 772 clones were sequenced and 684 high qualities EST were obtained which included 17 contings and 562 singlets. The redundancy was 7.41%, and the ratio of full-length cDNAs was around 67%. Annotation by the online blast go soften revealed that the most of genes were related to catalytic activity, binding, transcription regulator activity, transporter activity and Eenzyme regulator activity. The analysis result indicated the library was efficient and reliable. After further analysis, some new genes related with bitter gourd fruit ripening and softening were obtained.