甜菜基因BvPCNA-like的克隆及结构分析  

Isolation and Genomic-structure Analysis of BvPCNA-like Gene in Beta vulgaris(Sugar Beet)

作　　者：刘昕彤 申子萌 张琦[1,3,4] 张西宁 陶艳昵 李志烨 王希 赵春雷 LIU Xintong;SHEN Zimeng;ZHANG Qi;ZHANG Xining;TAO Yanni;LI Zhiye;WANG Xi;ZHAO Chunlei(College of Advanced Agriculture and Ecological Environment of Heilongjiang University/Sugar Beet Research Institute of Chinese Academy of Agricultural Sciences,Harbin 150080;College of Life Sciences and Technology,Longdong University,Qingyang,Gansu 745000;Key Laboratory of Sugar Beet Genetics and Breeding,Heilongjiang Province Common College/Sugar Beet Engineering Research Center of Heilongjiang Province,Harbin 150080;National Sugar Crops Improvement Center/Key Laboratory of North Sugar Crop Resource and Utilization,Chinese Academy of Agricultural Sciences,Harbin 150080;Gansu Key Laboratory of Protection and Utilization for Biological Resources and Ecological Restoration,Qingyang,Gansu 745000;China Blue Sky Ecological Technology(Beijing)Co.,Ltd.,Beijing 100083)

机构地区：[1]黑龙江大学现代农业与生态环境学院/中国农业科学院甜菜研究所,哈尔滨150080 [2]陇东学院生命科学与技术学院,甘肃庆阳745000 [3]黑龙江省普通高等学校甜菜遗传育种重点实验室/黑龙江省甜菜工程技术研究中心,哈尔滨150080 [4]国家糖料改良中心/中国农业科学院北方糖料作物资源与利用重点开放实验室,哈尔滨150080 [5]甘肃省高校陇东生物资源保护与利用省级重点实验室,甘肃庆阳745000 [6]中资蓝天生态科技(北京)有限公司,北京100083

出　　处：《中国糖料》2023年第2期16-25,共10页Sugar Crops of China

基　　金：博士后研究人员落户黑龙江科研启动资助金“甜菜分子标记开发与重要性状相关标记的筛选”(LBH-Q18108);2017年度黑龙江省省属高等学校基本科研业务费科技创新类重点项目“甜菜分子标记位点库的深度挖掘与候选SSR标记开发”(KJCXZD201713);财政部和农业农村部国家现代农业产业技术体系(糖料)建设项目“甜菜种质资源收集与评价”(CARS-170102)资助。

摘　　要：本文旨在利用分子标记位点克隆糖用甜菜(Beta vulgaris L.)中的BvPCNA-like基因的基因组位点及完整的编码区(Coding sequence,CDS),并对该CDS在基因组中的结构进行分析与验证,为进一步研究该基因的功能奠定基础。利用甜菜EST-SSR标记BvRE049,以糖用甜菜种质‘DP02’(标记检测阳性)为材料,通过电子克隆策略获得标记相关基因的基因组位点,进而克隆基因CDS片段;预测CDS在基因组中的外显子和内含子个数及内含子剪接位点分布范围,并设计PCR引物组合对以上结构进行验证;同时用在线生物信息学分析软件分析该基因产物的基本特征。克隆获得了甜菜基因BvPCNA-like的编码区,全长1164 bp,编码产物含387个氨基酸,分子量约42736.25 Da,等电点约4.46,在基因组中,该基因由3个内含子分隔成4个外显子。本研究克隆了BvPCNA-like基因的完整编码区,预测并验证了其在基因组位点中的结构,为进一步深入研究BvPCNA-like基因在甜菜体内的功能奠定了基础。To acquire the BvPCNA-like gene from Beta vulgaris(sugar beet)using a molecular marker locus,including both genome locus sequence and coding sequence(CDS),and to analyze and verify the structure of the CDS region in the genomic locus,so as to laying a foundation for the further functional research of this gene in the future.By virtue of an SSR molecular marker,BvRE049,and a sugar beet germplasm,DP02,which had been proved positive at the BvRE049 marker locus,the genome locus of the marker-related gene was isolated by in silico isolation strategy firstly,and then the coding sequence,CDS,was also isolated.CDS structure in the genomic locus was predicted in silico at first,including the number and distribution of exons and introns and regions of splicing sites,and then validated by PCR with a group of targeted designed primers.Meanwhile,online bioinformatics tools were used to analyze the general characteristics of the gene product.The target gene CDS,BvPCNA-like,was isolated with a length of 1164 bp.The coding product consists of 387 amino acids,whose molecular weight is about 42736.25 Da and isoelectric point is around 4.46.In the genome,the gene is separated into four exons by three introns.This study isolated the complete CDS of BvPCNA-like gene,predicted and verified its structure in the genome locus,so that laid a foundation for further studies on the function of BvPCNA-like gene in sugar beet.

关 键 词：甜菜 增殖细胞核抗原 基因克隆 编码区结构 生物信息学 

分 类 号：S566.3[农业科学—作物学]