CAJ | 학술논문

病毒诱导的基因沉默(virus induced gene silencing, VIGS)技术是近年来发展起来的一种反向遗传学快速研究基因功能的方法,对于植物特别是难于转化的大豆而言尤其适用.本研究采用PCR技术从大豆(Glycine max)基因组中克隆了八氢番茄红素去饱和酶(phytoene desaturase,PDS)基因的部分序列,命名为GmPDS(Glycine max PDS).该片段长430 bp,序列分析表明,该基因与大豆八氢番茄红素去饱和酶(GenBank 登录号:M64704)cDNA序列同源性为99%.双酶切GmPDS片段和烟草脆裂病毒载体(pTRV2),构建了重组载体pTRV2-GmPDS,并将该重组载体分别转化农杆菌C58C1/pMP90、GV3101和LBA4404,为分析pTRV载体是否可以浸染大豆奠定了基础.
병독유도적기인침묵(virus induced gene silencing, VIGS)기술시근년래발전기래적일충반향유전학쾌속연구기인공능적방법,대우식물특별시난우전화적대두이언우기괄용.본연구채용PCR기술종대두(Glycine max)기인조중극륭료팔경번가홍소거포화매(phytoene desaturase,PDS)기인적부분서렬,명명위GmPDS(Glycine max PDS).해편단장430 bp,서렬분석표명,해기인여대두팔경번가홍소거포화매(GenBank 등록호:M64704)cDNA서렬동원성위99%.쌍매절GmPDS편단화연초취렬병독재체(pTRV2),구건료중조재체pTRV2-GmPDS,병장해중조재체분별전화농간균C58C1/pMP90、GV3101화LBA4404,위분석pTRV재체시부가이침염대두전정료기출.
Virus induced gene silencing (VIGS) is increasingly being used as a reverse genetics tool to study functions of specific plant genes in a short time. It is especially useful for plants, especially soybean that is recalcitrant to transformation. A partial sequence of phytoene desaturase (PDS)was cloned from Glycine max using PCR method and termed as GmPDS (Glycine max PDS). GmPDS indicates 430 bp length,and exhibits 99% identity to the cDNA sequences that PDS was obtained from GenBank database (GenBank accession number M64704) at nucleotide acids level.The GmPDS and tobacco rattle virus vector (pTRV2) were digested by double restriction enzymes and recombinant pTRV2-GmPDS vector was constructed,which was transformed into Agrobacterium C58C1/pMP90, GV3101and LBA4404, respectively.