水稻谷蛋白基因GluB-6的cDNA克隆及表达

cDNA Cloning and Expression Analysis of GluB-6 in Rice

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eDNA序列全长为1517bp，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。

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 eDNA, named GluB-6（GenBank accession number is AY429651）, was cloned. DNA sequencing analysis and in vitro expression results showed that the cloned eDNA 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.