紫花苜蓿MsSQE1的克隆及对皂甙合成的功能分析

康俊梅,张俏燕,蒋旭,王珍,张铁军,龙瑞才,崔会婷,杨青川

Cloning MsSQE1 from Alfalfa and Functional Analysis in Saponin Synthesis

JunMei KANG,QiaoYan ZHANG,Xu JIANG,Zhen WANG,TieJun ZHANG,RuiCai LONG,HuiTing CUI,QingChuan YANG

表1 本研究所用引物序列

Table 1 The sequences of primers used in this study

引物名称 Primer name 引物序列 Sequence of primer (5′-3′) 用途 Function P1 GTTCTTTGAGATGGATTATCAGTAT ORF分离 ORF isolation P2 AAATGACATGAATACTACAGTGACA SQE1-PE-SPt-f GAAAAGAAGAAAACAGGTTCAAGTT 原核表达 Prokaryotic expression SQE1-PE-f ATGGATTATCAGTATATTCTTGGAGGG SQE1-PE-r ATGGACAGGAGGCATTCTGTAATAT SQE1-GFP-f CCctcgagATGGATTATCAGTATATTCTTGGAGGG 亚细胞定位 Subcellular localization SQE1-GFP-r TGgtcgacATGGACAGGAGGCATTCTGTAATAT SQE1-RTf-1 GCAAAGGCTCCTCTCACCATAG 实时荧光定量 PCR/qRT-PCR SQE1-RTr-1 GCACATCAACCAAACAGCGAAT Actin-s CAAAAGATGGCAGATGCTGAGGAT 内参基因 House-keeping gene Actin-a CATGACACCAGTATGACGAGGTCG SQE1-P1 tctagaATGGATTATCAGTATATTCTTGGAGGG 扩增目的基因 Amplification of target genes SQE1-P2 ggatccTTAATGGACAGGAGGCATTCTGTAATATGC 35S-f ACTATCCTTCGCAAGACCCTTCCTC 转基因株系的鉴定 Identification of transgenic lines SQE1-r TTAATGGACAGGAGGCATTCTGTAAT