CAJ | 학술논문

濒危植物海南粗榧富含多种抗癌活性次生代谢产物.为研究这些产物的生物代谢途径和表达调控,通过同源克隆和RACE技术获得了海南粗榧紫杉醇生物合成途径中通用前体牻牛儿基牻牛儿焦磷酸合成酶(GGPPs)基因全长并进行了生物信息学分析.该基因命名为CmGGPPs (GenBank登录号:JX971119),并进一步利用不同激发子诱导研究了该基因对不同逆境信号的响应情况.结果表明:海南粗榧GGPPs基因cDNA全长1694 bp,包含一个1182 bp的ORF框,编码393个氨基酸;预测该基因编码蛋白质分子量为42.91 kD,其等电位为7.12.通过BLAST比对结果分析,海南粗榧GGPPs基因全长核苷酸序列与红豆杉科植物的同源性最高可达91%,氨基酸序列有89%的相似性.同时,GGPPs基因氨基酸序列同其他植物也有较高的同源性.用不同类型激发子处理发现,GGPPs在诱导后表达量均上升,但不同激发子诱导的基因表达强度和速率存在明显区别.
빈위식물해남조비부함다충항암활성차생대사산물.위연구저사산물적생물대사도경화표체조공,통과동원극륭화RACE기술획득료해남조비자삼순생물합성도경중통용전체방우인기방우인초린산합성매(GGPPs)기인전장병진행료생물신식학분석.해기인명명위CmGGPPs (GenBank등록호:JX971119),병진일보이용불동격발자유도연구료해기인대불동역경신호적향응정황.결과표명:해남조비GGPPs기인cDNA전장1694 bp,포함일개1182 bp적ORF광,편마393개안기산;예측해기인편마단백질분자량위42.91 kD,기등전위위7.12.통과BLAST비대결과분석,해남조비GGPPs기인전장핵감산서렬여홍두삼과식물적동원성최고가체91%,안기산서렬유89%적상사성.동시,GGPPs기인안기산서렬동기타식물야유교고적동원성.용불동류형격발자처리발현,GGPPs재유도후표체량균상승,단불동격발자유도적기인표체강도화속솔존재명현구별.
@@@@Endangered plant, Cephalotaxus mannii, is known as containing rich and various anti-cancer active secondary metabolites. In order to study the metabolitic pathway and expression regulation, the geranyl geranyl diphosphate synthase (GGPPs) gene that catalyzes and synthesizes the common precursor in paclitaxel biosynthetic pathway, was cloned from Cephalotaxus mannii by means of homologous cloning approach and RACE technique. The cloned gene was names as CmGGPPs and deposited in GenBank with Accession number JX971119. The sequence analysis showed that the cDNA sequence had 1 694 bp in length with a poly A tail, which contains a 1 182 bp of open reading frame (ORF) encoding a 393 amino acid residuals. Bioinformatics analysis predicted that it encodes a 42.91 kD predicted protein with isoelectric point 7.12. Blast results indicated that the cloned gene share 91%sequence homology as well as 89%amino acid similarity with the reported GGPPs of other plants in the family of Taxaceae Meanwhile, GGPPs gene in Amino acid sequences also shares high homology with other plants. Through the treatment with different types of elicitors, the results showed that GGPPs expression levels increased after elicitor's induction, but there were distinct different strength and rate of the gene expression in three elicitor's treatments.