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吉林省自然科学基金(20101568)

作品数:2 被引量:7H指数:1
相关作者:郭庆勋李彦磊贾承国阮文渊刘羽更多>>
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辣椒中辣椒素生物合成途径基因Kas克隆及表达研究被引量:7
2011年
3-氧酰基[酰基载体蛋白]还原酶(3-oxoacyl-[acyl-carrier-protein]synthase,Kas)在辣椒素生物合成途径中起着重要作用,但其全长DNA序列仍未见报道。以辣椒益都红基因组为模板,根据同源性设计特异引物PCR扩增得到辣椒Kas基因DNA序列(GenBank登录号:HQ229922)。Kas DNA序列全长3 456 bp,有8个外显子区域,编码488个氨基酸;推测蛋白分子质量为52.2 ku,等电点8.24。经同源比对分析得出,辣椒Kas与其他物种Kas有一定同源性;其中,与小米椒的遗传距离为0.0723。通过半定量RT-PCR分析发现,Kas在胎座中的表达量要高于肉质体中的表达量,授粉10 d后Kas的表达量逐渐降低。另外,植物激素油菜素内酯(Brassinosteroid,BR)对辣椒胎座Kas基因的表达有促进作用,在处理24 h后Kas表达量达到最高,随后逐渐降低。该基因的克隆与表达分析为深入研究辣椒Kas基因表达调控和辣椒素的生物合成机制奠定了重要基础。
阮文渊贾承国李彦磊郭庆勋
关键词:辣椒素油菜素内酯
Cloning, Expression and Analysis of pal, a Positive Gene Involved in Capsaicin Biosynthesis in Capsicum chinense Jacq被引量:1
2013年
The cDNA sequence of Capal gene was cloned from Capsicum chinense Jacq by RT-PCR and sequenced. Bioinformatics analysis showed that Capal en- codes a putative polypeptide of 683 amino acids with a calculated molecular mass of 74.2 kD and a theoretical pl of 6.9. Multiple sequence alignments and phyloge- netic tree analyses showed that Capal protein of C. chinense is similar to that of Capsicum annuum var. conoides, with an overall sequence similarity of 96%. The prokaryotic expression vector pET-32a-pal was constructed and induced to express in E. coil BL21. The SDS-PAGE analysis showed that the relative molecular mass of the induced new protein is about 74 kD, which was basically identical with that predicted by DNAMAN software (74.3 kD), Real-time PCR analysis showed that ex- ogenous jasmonic acid (JA) promoted pal expression. The accumulation of capsaicin in pepper was analyzed by high performance liquid chromatography (HPLC), and the results indicated that exogenous jasmonic acid (JA) can promote the synthesis of capsaicin. This study will provide valuable experimental basis for the research of transcription regulation and explaining the gene function of pal.
刘艳梅杨帆段军涛刘羽王晶莹郭庆勋
关键词:CAPSAICIN
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