论文部分内容阅读
为分析洋葱AcAG基因(GenBank ID:JX974431)的功能,本研究根据课题组前期获得的洋葱C类MADS-box基因AcAG的cDNA序列设计特异引物,利用RT-PCR技术从洋葱花瓣中分离到了AcAG基因的全长CDS序列,连接到植物载体pCAMBIA1304中,构建超表达载体pCAMBIA1304-AcAG(+)。采用冻融法将重组植物表达载体转入到农杆菌菌株LBA4404中,并利用花序侵染法对野生型拟南芥进行遗传转化。经RT-PCR和酶切鉴定,结果表明,AcAG过表达载体构建成功;转基因植株基因组PCR和RT-PCR分析结果表明,外源基因AcAG已经整合到拟南芥基因组中,并在转录水平上发生表达。
In order to analyze the function of Onion AcAG gene (GenBank ID: JX974431), we designed specific primers based on the cDNA sequence of onion C class MADS-box gene AcAG previously obtained from the onion. The AcAG gene was isolated from onion petals by RT-PCR The full-length CDS sequence was ligated into the plant vector pCAMBIA1304 to construct the overexpression vector pCAMBIA1304-AcAG (+). The recombinant plant expression vector was transformed into Agrobacterium strain LBA4404 by freeze-thaw method, and the wild-type Arabidopsis thaliana was transformed by inflorescence. The results of RT-PCR and restriction enzyme digestion showed that AcAG overexpression vector was successfully constructed. The results of PCR and RT-PCR showed that AcAG gene was integrated into Arabidopsis genome and transcribed at the transcriptional level expression.