Chinese Journal of Rice Science

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cDNA Cloning and Expression Analysis of GluB-6 in Rice

NIU Hong-bin , WANG Yi-hua , ZHAI Hu-qu , WAN Jian-min   

  • Received:1900-01-01 Revised:1900-01-01 Online:2007-03-10 Published:2007-03-10

水稻谷蛋白基因GluB-6的cDNA克隆及表达

牛洪斌1;王益华1;翟虎渠2;万建民1,2,*   

  1. 1南京农业大学 作物遗传与种质创新国家重点实验室, 江苏 南京 210095; 2中国农业科学院 作物科学研究所, 北京100081; *通讯联系人, E-mail: wanjm@caas.net.cn

Abstract: Based on the conserved amino acid sequences of rice glutelin gene family, one highly homologous BAC clone sequence from the rice genome database of GenBank was obtained and a putative glutelin gene was predicted. After amplified by RT-PCR with the specific primers designed on the basis of predicted nucleotide sequence, a new glutelin gene cDNA, named GluB-6(GenBank accession number is AY429651), was cloned. DNA sequencing analysis and in vitro expression results showed that the cloned cDNA was 1517 bp, and carried entire coding sequences,which encoded a 495 amino acid protein; this corresponded to the size of the glutelin protein family. Homology analysis showed that the deduced amino acid sequence of GluB-6 shared 53.6%-82.8% identity with rice glutelin gene family other else. Northern blot analysis indicated that GluB-6 expressed especially in rice endosperm.

Key words: rice, glutelin gene, cloning, expression

摘要: 根据已克隆谷蛋白基因的保守氨基酸序列搜索基因组数据库,获得与之高度同源的水稻基因组序列,通过生物软件进行基因预测和验证,用RTPCR法克隆得到1个新的谷蛋白基因的cDNA克隆。核酸序列分析和体外表达结果表明,该基因cDNA序列全长为1517 bp,含有1个编码495个氨基酸残基的开放阅读框(ORF),推导的氨基酸序列与谷蛋白基因家族的相似性介于53.6%~82.8%,并与B亚族谷蛋白基因的同源性更高,因此命名为GluB6(GenBank注册号AY429651)。Northern杂交显示,GluB6基因具有高度的胚乳表达特性。

关键词: 水稻, 谷蛋白基因, 克隆, 表达