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当前位置: 首页 > 文章 > 拟南芥高亲和性K~+载体蛋白基因cDNA克隆及其序列特征分析 草业学报 2013,22 (6) 230-238
Position: Home > Articles > Cloning and analysis of sequence characteristics of cDNA encoding the AtHKT1 gene from Arabidopsis thaliana leaves Acta Prataculturae Sinica 2013,22 (6) 230-238

拟南芥高亲和性K~+载体蛋白基因cDNA克隆及其序列特征分析

作  者：
王丽;张俊莲;张金文;刘玉汇;白江平;余斌;杨宏羽;王蒂
单  位：
甘肃省作物遗传改良与种质创新重点实验室
关键词：
拟南芥;高亲和性K+载体蛋白;RT-PCR技术;生物信息学分析
摘  要：
以NaCl胁迫处理的拟南芥幼苗叶片为材料,用RNA提取试剂盒抽提总RNA,通过RT-PCR技术和DNA序列测定分析,证实获得了拟南芥高亲和性K+载体蛋白基因(AtHKT1)的cDNA序列。该cDNA全长1521bp,包括506个氨基酸和1个终止密码子序列,且与原序列(accession number AF237672)同源性为99.34%,但与其他科植物HKT1基因同源性较低,注册该基因到GenBank中,注册号为AY685182。利用生物信息学相关软件分析预测AtHKT1基因蛋白质功能和结构,结果发现,该蛋白分子量为57.45kD,理论等电点为9.33;氨基酸序列中第1～40个氨基酸属信号肽序列;第152～500个氨基酸属Trk H阳离子转运体蛋白保守结构域,并存在蛋白激酶C,酪氨酸蛋白激酶,依赖cAMP/cGMP蛋白激酶磷酸化,糖基化和豆蔻酰化等功能位点;该基因编码的蛋白有10个跨膜结构,N末端、C末端及中部等多个跨膜区具疏水性,符合载体类运输蛋白特点。表明本研究获得了拟南芥AtHKT1基因。
译  名：
Cloning and analysis of sequence characteristics of cDNA encoding the AtHKT1 gene from Arabidopsis thaliana leaves
作  者：
WANG Li;ZHANG Jun-lian;ZHANG Jin-wen;LIU Yu-hui;Bai Jiang-ping;YU Bin;YANG Hong-yu;WANG Di;Gansu Key Laboratory of Crop Genetic & Germplasm Enhancement;College of Life Sciences and Technology,Gansu Agricultural University;Gansu Provincial Key Lab of Aridland Crop Science;College of Agronomic,Gansu Agricultural University;
关键词：
Arabidopsis thaliana;;high-affinity potassium transporter;;RT-PCR;;bioinformatic analysis
摘  要：
The full length coding sequence of AtHKT1was cloned via RT-PCR(reverse transcription-polymerase chain reaction)from the leaves of Arabidopsis thaliana seedling under NaCl stress.The full-length cDNA contained 1521bp and encoded a polypeptide of 506amino acids and a termination codon.The sequence shared 99.34% homology with the reported sequence(accession number AF237672),but the homologies were very low compared with similar genes from other plants.The gene was submitted to GenBank and was given the registration number AY685182.Bioinformatics softwares were used to predict and analyze the protein function and structure of the AtHKT1gene cDNA sequence.The molecular weight of the deduced polypeptide of AtHKT1 was 57.45kD.The theoretical pI was 9.33.The N-terminal amino acids from 1to 40formed a classic signal peptide sequence.The peptide from 152to 500was a Trk H cation transporter conserved domain.It might also carry protein kinase C,tyrosine kinase,cAMP-and cGMP-dependent protein kinase phosphorylation,glycosylation,N-myristoylation sites and other function sites.The deduced peptide contained 10transmembrane domains.The transmembrane domains near the N-terminus,the C-terminus and the middle part of the peptide displayed high hydrophobicity.Therefore,it is suggested that the gene product of AtHKT1might play roles in facilitating cation transportation.

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