摘要
初探StTCP13基因在盐胁迫方面的功能,为StTCP13在马铃薯非生物逆境响应的功能研究奠定基础。采用同源克隆策略获得StTCP13基因编码区全长,利用生物信息学分析蛋白结构域并构建系统进化树,qRT-PCR技术分析该基因表达模式,双酶切法构建StTCP13-pGEX6P1原核表达载体,利用大肠杆菌BL21表达StTCP13-GST标签融合蛋白,观察转StTCP13大肠杆菌BL21在盐胁迫培养基上的表型。结果显示,从马铃薯品种费乌瑞它中克隆得到了StTCP13基因,成功构建了StTCP13-GST标签融合蛋白的pGEX-6P-1原核表达载体,并在大肠杆菌BL21中诱导表达了目的蛋白。生物信息学分析结果表明,该基因CDS全长669 bp,编码一个由222个氨基酸组成的蛋白质,该蛋白质含有1个保守的TCP结构域,融合蛋白大小约为51 ku。系统进化树结果显示,马铃薯StTCP13与潘那利番茄SpTCP13、甜椒CaTCP13亲缘关系较近。表达分析结果显示,该基因在根中表达量最高,在芽眼中表达量最低,其表达受高盐胁迫诱导。盐胁迫表型分析结果表明,StTCP13能提高大肠杆菌对高盐胁迫的耐受性。综上,说明StTCP13在响应盐胁迫逆境中发挥一定功能。
In order to investigate the function of StTCP13 in salt stress and lay the foundation for the functional study of StTCP13 in response to abiotic stress of potato.The coding sequence of StTCP13 was obtained by homologous cloning.The protein domain and phylogenetic tree were analyzed by bioinformatics.The results showed that the StTCP13 gene was cloned from the potato variety Favorita.The coding sequence length of StTCP13 was 669 bp,encoding a protein composed of 222 amino acid residues.The results of prokaryotic expression suggested that the molecular weight of fusion protein was 51 ku.Protein domain analysis suggested that the protein contained a conserved TCP domain.The results of phylogenetic tree revealed that the protein had a high homology with the TCP13 in Solanum pennellii and Capsicum annuum.Expression pattern of StTCP13 was analyzed by qRT-PCR.Expression analysis showed that the StTCP13 exhibited different expression levels in root,stem,leaf,bud and bud eye in potato.The StTCP13 had high expression level in root,low expression level in bud eye,in addition,the FPKM of StTCP13 in Spud DB showed that the expression of StTCP13 was induced by high-salinity,drought and ABA treatments.The StTCP13-pGEX6P1 prokaryotic expression vector was constructed by double enzyme digestion.Prokaryotic expression results showed that the StTCP13-GST tag fusion protein was successfully expressed in Escherichia coli BL21.Salt stress phenotype analysis suggested that StTCP13 could improve the tolerance to salt stress in E.coli BL21.Above all,the StTCP13 might play a role in response to salt stress in potato.
作者
李佳皓
谢敏秋
万传银
左瑞洁
鲁黎明
李立芹
LI Jiahao;XIE Minqiu;WAN Chuanyin;ZUO Ruijie;LU Liming;LI Liqin(College of Agronomy, Sichuan Agricultural University, Chengdu 611130,China;National Demonstration Center for Experimental Crop Science Education, Chengdu 611130,China)
出处
《华北农学报》
CSCD
北大核心
2021年第2期33-39,共7页
Acta Agriculturae Boreali-Sinica
基金
四川省科学技术厅项目(3742018JY0078)。
关键词
马铃薯
盐胁迫
表型分析
TCP
表达模式
原核表达
转录因子
Potato
Salt stress
Phenotypic analysis
TCP
Expression pattern
Prokaryotic expression
Transcription factor
