CAJ | 학술논문

根据MaizeDGB数据库中ZmCIPK6基因序列设计引物, 采用RT-PCR技术从玉米中克隆了1个ZmCIPK6基因. ZmCIPK6基因cDNA全长1 739 bp,5'-非编码区长281 bp, 3'-非编码区长111 bp, 编码区长1 347 bp,编码448个氨基酸,预测分子量为48.8 KDa,等电点为9.14. 氨基酸同源性分析发现, ZmCIPK6与高粱SbCIPK6同源性较高. 根据ZmCIPK6的基因序列和植物表达载体pCAMBIA3301的多克隆位点设计带有限制性内切酶位点的特异性引物,以质粒pMD18-T-ZmCIPK6为模板,PCR扩增Zm-CIPK6基因片段,双酶切目的片段及载体,回收后连接,构建成该基因的植物表达载体.经PCR检测及测序鉴定,表明成功构建了ZmCIPK6基因的植物表达载体,为进一步遗传转化研究基因的功能奠定基础.
근거MaizeDGB수거고중ZmCIPK6기인서렬설계인물, 채용RT-PCR기술종옥미중극륭료1개ZmCIPK6기인. ZmCIPK6기인cDNA전장1 739 bp,5'-비편마구장281 bp, 3'-비편마구장111 bp, 편마구장1 347 bp,편마448개안기산,예측분자량위48.8 KDa,등전점위9.14. 안기산동원성분석발현, ZmCIPK6여고량SbCIPK6동원성교고. 근거ZmCIPK6적기인서렬화식물표체재체pCAMBIA3301적다극륭위점설계대유한제성내절매위점적특이성인물,이질립pMD18-T-ZmCIPK6위모판,PCR확증Zm-CIPK6기인편단,쌍매절목적편단급재체,회수후련접,구건성해기인적식물표체재체.경PCR검측급측서감정,표명성공구건료ZmCIPK6기인적식물표체재체,위진일보유전전화연구기인적공능전정기출.
According to the sequence of ZmCIPK6 in MaizeDGB database,primers were designed. RT-PCR method was used to clone ZmCIPK6 gene. A gene coding for ZmCIPK6 was isolated from maize (Zea mays).The full length ZmCIPK6 cDNA was 1 739 bp,including a 281 bp 5'-UTR,an ORF of 1 347 bp,and a 111 bp 3'-UTR.This cDNA sequence encoded a polypepi-de of 448 amino acid residues with a predicted molecular mass of 48.8 kDa and a basic isoelectric point of 9.14. The deduced amino acid sequence had a high homology with SbCIPK6 from Sorghum bicolor.According to the restriction enzyme sites of expression vector pCAMBIA3301 and sequence of ZmCIPK6 gene,one pair of primers containing restriction enzyme sites were designed. The ZmCIPK6 gene was obtained from pMD18-T-ZmCIPK6 by PCR. PCR product and the plasmid pCAMBIA3301 were digested by the corresponding restricted enzymes respectively,and then the ZmCIPK6 gene was cloned into pCAMBI-A3301 vector.PCR and sequencing results suggest that plant expression vector of ZmCIPK6 gene was constructed successfully.