CAJ | 학술논문

[目的]获得草莓磷脂酶Dα (PLDα)的HKD2功能区基因序列构建反义植物表达载体.研究PLDα基因对草莓果实耐贮性的影响.[方法]以草莓完熟果实为材料提取总RNA,利用RT-PCR技术扩增得到PLDα基因的cDNA片段,将其T-A克隆至pMD19-T simple vector上,测序正确后克隆载体经双酶切消化,目的片断反向插入到植物表达载体pBI121中.[结果]构建草莓PLDα基因的反义植物表达载体pBI121-PLD.[结论]通过冻融法将携带反义cDNA的植物表达载体质粒导入根癌农杆菌LBA44404,为进一步通过反义技术延长草莓果实贮藏的寿命奠定了基础.
[목적]획득초매린지매Dα (PLDα)적HKD2공능구기인서렬구건반의식물표체재체.연구PLDα기인대초매과실내저성적영향.[방법]이초매완숙과실위재료제취총RNA,이용RT-PCR기술확증득도PLDα기인적cDNA편단,장기T-A극륭지pMD19-T simple vector상,측서정학후극륭재체경쌍매절소화,목적편단반향삽입도식물표체재체pBI121중.[결과]구건초매PLDα기인적반의식물표체재체pBI121-PLD.[결론]통과동융법장휴대반의cDNA적식물표체재체질립도입근암농간균LBA44404,위진일보통과반의기술연장초매과실저장적수명전정료기출.
[Objective]The research aimed to obtain HKD2 domain in PLD gene and construct its antisense plant expression vector,so as to study the effect of PLDα gene on the storage property,[Method]total RNA was extracted from the mature fruit of strawberry,then cDNA was amplified by reverse transcriptase polymerase chain reaction(RT-PCR) and to clone T-A into pMD19-T simple vector. The positive clones were sent to sequence. [Result]After confirming its validity,the PLDα gene fragment was inverted into plant expression vector pBI121 to construct antisense expression vector pBI121-PLD.[Conclusion]Then the recombinant plasmid was transferred into Agrobacterium tumerfaciens LBA4404 by freeze-thaw method to further lay the base for extending the strawberry's storage.