本文關(guān)鍵詞： 芥藍(lán) 八氫番茄紅素脫氫酶 基因克隆 原核表達(dá)　出處：《基因組學(xué)與應(yīng)用生物學(xué)》2016年07期 　論文類型：期刊論文

【摘要】：本研究以‘明豐香菇’芥藍(lán)葉片總RNA為模板,采用同源克隆法設(shè)計并合成特異性引物,采用反轉(zhuǎn)錄PCR技術(shù)克隆得到芥藍(lán)類胡蘿卜素生物合成關(guān)鍵結(jié)構(gòu)基因-八氫番茄紅素脫氫酶基因Ba PDS1。序列分析表明,該基因c DNA序列長1 692 bp,編碼563個氨基酸,推測蛋白等電點為5.96,理論分子量為63.15 k D;序列比對發(fā)現(xiàn)芥藍(lán)Ba PDS1基因編碼的氨基酸序列與甘藍(lán)、花椰菜、油菜等植物的PDS具有很高的同源性;系統(tǒng)進化樹分析表明,芥藍(lán)PDS1與甘藍(lán)PDS1蛋白的親緣關(guān)系最為接近。之后構(gòu)建p EASY-Blunt E1-Ba PDS1原核表達(dá)載體,轉(zhuǎn)入Transetta(DE3)感受態(tài)細(xì)胞中,誘導(dǎo)后獲得特異表達(dá)蛋白條帶,且大部分以包涵體形式存在。本研究為揭示芥藍(lán)Ba PDS1基因功能提供了一定的理論依據(jù)。
[Abstract]:In this study, the specific primers were designed and synthesized by homologous cloning using the total RNA of 'Mingfeng Lentinus edodes' leaves as template. The key structural gene of carotenoid biosynthesis of kale was cloned by reverse transcription PCR. The gene Ba PDS1 of lycopene dehydrogenase of octahydrolycopene was sequenced. The c DNA sequence is 1 692bpand encodes 563 amino acids. The deduced isoelectric point of the protein is 5.96 and the theoretical molecular weight is 63.15kD; Sequence alignment showed that the amino acid sequence encoded by Ba PDS1 gene of kale had high homology with PDS of cabbage, cauliflower, rape and so on. Phylogenetic tree analysis showed that the relationship between kale PDS1 and cabbage PDS1 protein was the most close, and then the prokaryotic expression vector of p EASY-Blunt E1-Ba PDS1 was constructed. Transfered into transettade3) cells, the specific expression protein bands were obtained after induction. Most of them exist in the form of inclusion bodies. This study provides a theoretical basis for revealing the function of Ba PDS1 gene in kale.
【作者單位】： 四川農(nóng)業(yè)大學(xué)園藝學(xué)院;
【基金】：國家自然科學(xué)基金項目(31500247) 四川省教育廳重點項目(14ZA0016) 四川農(nóng)業(yè)大學(xué)本科生科研興趣培養(yǎng)計劃項目(2015009)共同資助
【分類號】：S635.9
【正文快照】： Cloning and Prokaryotic Expression of Ba PDS1 Gene in Brassica alboglabraXue Shengling Zhang Fen Xia Xue Leng Zhenmin Chen Qing Tang Haoru Sun Bo*College of Horticulture,Sichuan Agricultural University,Chengdu,611130*Corresponding author,14099@sicau.edu.，

本文編號：1487906