两个烟草多聚半乳糖醛酸酶抑制蛋白基因的克隆和表达分析

doi:10.7668/hbnxb.2015.01.006

华北农学报 ›› 2015, Vol. 30 ›› Issue (1): 29-34. doi: 10.7668/hbnxb.2015.01.006

所属专题： 烟草；生物技术；

两个烟草多聚半乳糖醛酸酶抑制蛋白基因的克隆和表达分析

林世锋¹, 安雪琴², 杨承², 王仁刚¹, 任学良¹, 赵杰宏¹, 付强¹

1. 贵州省烟草科学研究院, 烟草行业分子遗传重点实验室, 贵州贵阳 550081;
2. 贵州省烟草公司铜仁市公司, 贵州铜仁 554300

收稿日期:2014-09-10 出版日期:2015-02-28
通讯作者: 付强(1984-),男,贵州赤水人,副研究员,博士,主要从事烟草遗传育种与蛋白质组学研究。
作者简介:林世锋(1978-),男,黑龙江牡丹江人,副研究员,博士,主要从事烟草遗传育种与分子生物学研究。
基金资助:
中国烟草总公司重点项目(110201302004);贵州省科学技术基金项目(黔科合J字[2012]2256号);贵州省优秀青年科技人才培养对象专项资金(黔科合人字【2013】02 号)

Cloning and Tissue Expression Analysis of Two Polygalacturonase Inhibiting Protein Genes from Tobacco

LIN Shi-feng¹, AN Xue-qin², YANG Cheng², WANG Ren-gang¹, REN Xue-liang¹, ZHAO Jie-hong¹, FU Qiang¹

1. Guizhou Academy of Tobacco Science, Key Laboratory of Molecular Genetics, CNTC, Guiyang 550081, China;
2. Tongren Tobacco Company of Guizhou Province, Tongren 554300, China

Received:2014-09-10 Published:2015-02-28

摘要/Abstract

摘要： 为了研究多聚半乳糖醛酸酶抑制蛋白(PGIP)在烟草中的生物学功能,运用生物信息学方法,结合RT-PCR和SMART RACE技术,从烟草中克隆到2个多聚半乳糖醛酸酶抑制蛋白(PGIP)基因cDNA和DNA全长序列,分别命名为 NtPGIP1 (GenBank登录号:KF317203)和 NtPGIP2 (GenBank登录号:KF317204)。NtPGIP1 基因全长为1 413 bp,编码338个氨基酸; NtPGIP2 基因全长为1 185 bp,编码329个氨基酸;两基因均没有内含子序列,核苷酸序列有50%的一致性,编码的氨基酸序列有54%的一致性。两基因编码蛋白均含有植物PGIP蛋白特有的重复保守序列LXXLXXLXXLXLXXNXLXGXIPXX。对PGIP基因在烟草不同组织表达量分析发现,两基因在根、茎、叶和芽中均有表达,其中均在茎中表达量最高,其次是根,叶中表达量很低。

关键词: 烟草, 多聚半乳糖醛酸酶抑制蛋白, 基因克隆, 组织表达

Abstract: In order to study the biological functions of tobacco PGIP, two full-length polygalacturonase inhibiting protein (PGIP) cDNA and DNA were cloned from Nicotiana tabacum by bioinformation, RT-PCR and SMART RACE technology, which were named NtPGIP1 (GenBank Number KF317203) and NtPGIP2 (GenBank Number KF317204), respectively.The full length NtPGIP1 was 1 413 bp which coding a protein for 338 amino acids.The full length NtPGIP2 was 1 185 bp which coding for 329 amino acids.Sequencing analysis showed that the identical amino acid sequence of the two genes was 54%, while the identical nucleotide sequence was 50%, and no intron sequences were found in the two genes.The amino acid sequences of the two proteins both contained plant PGIP specific conserved leucine repeat sequence LXXLXXLXXLXLXXNXLXGXIPXX.The gene expression patterns were analyzed by Real-time quantitative PCR.The results indicated that the expression of the two genes was detected in all tissues with the highest expression level in stems, followed in roots, and lowest level in leaves.

Key words: Nicotiana tabacum, Polygalacturonase inhibiting protein (PGIP), Gene cloning, Tissue expression

中图分类号:

S785
Q78

林世锋, 安雪琴, 杨承, 王仁刚, 任学良, 赵杰宏, 付强. 两个烟草多聚半乳糖醛酸酶抑制蛋白基因的克隆和表达分析[J]. 华北农学报, 2015, 30(1): 29-34. doi: 10.7668/hbnxb.2015.01.006.

LIN Shi-feng, AN Xue-qin, YANG Cheng, WANG Ren-gang, REN Xue-liang, ZHAO Jie-hong, FU Qiang. Cloning and Tissue Expression Analysis of Two Polygalacturonase Inhibiting Protein Genes from Tobacco[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(1): 29-34. doi: 10.7668/hbnxb.2015.01.006.

http://www.hbnxb.net/CN/Y2015/V30/I1/29

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两个烟草多聚半乳糖醛酸酶抑制蛋白基因的克隆和表达分析

Cloning and Tissue Expression Analysis of Two Polygalacturonase Inhibiting Protein Genes from Tobacco

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