中国水仙锌指蛋白NtPLATZ1的克隆与表达分析  

Cloning and expression analysis of NtPLATZ1 gene from Narcissus tazetta var. chinensis

作　　者：林江波[1] 王伟英[1] 李海明[1] 吴水金[1] 邹晖[1] 李跃森[1] 戴艺民[1] 

机构地区：[1]福建省农业科学院甘蔗研究所,福建漳州363005

出　　处：《西北农林科技大学学报（自然科学版）》2016年第10期165-170,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：福建省自然科学基金项目(2014J01098)

摘　　要：【目的】克隆中国水仙植物AT富集序列锌结合蛋白基因(NtPLATZ1),为研究该基因的生物学功能奠定基础。【方法】利用RACE技术克隆了NtPLATZ1cDNA全长,利用生物信息学方法分析其核苷酸及氨基酸序列,将NtPLATZ1与原核表达载体pGEX-4T-3连接,转化BL21并进行诱导表达,通过半定量PCR分析用多效唑处理后该基因在中国水仙不同生长时期叶片中的表达情况。【结果】NtPLATZ1的cDNA全长1 024bp,包含1个642bp的完整阅读框架,编码213个氨基酸;编码蛋白具有PLATZ superfamily蛋白保守区,与大豆(Glycine max,AII19314.1)PLATZ蛋白序列的同源性最高,为81%。原核表达结果表明,NtPLATZ1在大肠杆菌中能有效表达。半定量RT-PCR分析表明,喷雾多效唑后,NtPLATZ1基因在中国水仙不同生长时期叶片中的表达量先上升后下降,抽葶期叶片的表达量最高;喷雾清水后NtPLATZ1基因表达量先下降后又有所回升,抽葶期和始花期的表达量低于长叶期和盛花期。【结论】成功克隆了中国水仙NtPLATZ1基因,多效唑处理能明显诱导该基因的表达。【Objective】This study aimed to clone plant AT-rich sequence and zinc-binding protein gene from Narcissus tazetta var.chinensis（NtPLATZ1）for studying its biological function.【Method】RACE and RT-PCR technique were used to clone NtPLATZ1,the bioinformatics analysis was conducted to analyze its sequence,and semi-quantitative RT-PCR was used to detect its expression.NtPLATZ1 was inserted into pGEX-4T-3,the prokaryotic expression vector was transformed into BL21,and protein expression was induced.【Result】The full length of NtPLATZ1 cDNA was 1 024 bp with an 642 bp ORF,which encoded a protein of 213 amino acids with a conservative domain of PLATZ superfamily.NtPLATZ1shared81%identity with PLATZ of Glycine max（AII19314.1）.NtPLATZ1 was predicted an unstable protein.Prokaryotic expression showed that recombinant pGEX-NtPLATZ1 had high expression in E.coli BL21.Semi-quantitative RT-PCR revealed that NtPLATZ1 expression was enhanced at scape stage and decreased hereafter after it was sprayed with paclobutrazol,while it was decreased at scape and early flowering stages without paclobutrazol.【Conclusion】NtPLATZ1was clone and its expression was enhanced significantly by paclobutrazol.

关 键 词：中国水仙 PLATZ 基因表达 锌指蛋白 

分 类 号：Q78[生物学—分子生物学] S682.21[农业科学—观赏园艺]