昆虫学报2011，Vol.54Issue(7)：746-753,8.
家蚕Aly/REF的基因克隆、序列分析及其细胞定位
Cloning, sequence analysis and cellular localization of Aly/REF from the silkworm, Bombyx mori
摘要
Abstract
RNA and export factor binding proteins (Aly/REF) play important roles in RNA stability, processing and nuclear export. Referring to the sequence of Bombyx mori aly gene ( GenBank accession no. DQ497195.1 ), the aly/refgene (Bmaly/ref) from the silkworm, B. mori, was cloned via RT-PCR. The sequencing result showed that the open reading frame (ORF) of Bmaly/ref is 765 bp in length, encoding 254 amino acid residues, and the amino acid sequence of BmAIy/REF showed 49.7％ and 52.7％ identities with the homologs of Drosophila melanogaster and Mus musculus, respectively. The structure prediction displayed that BmAIy/REF contains RNA binding domain (RRM), REF-N and REF-C motif of REF subfamily. Phylogenetic analysis showed that the insect Aly/REF proteins clustered to a group and the BmAly/REF was close to Aly/REF proteins of Apis mellifera and Tribolium castaneum. The BmAly gene was inserted into expression vector pGS21 a ( + ), and then the recombinant protein was expressed in Escherichia coli and used to immunize mice to prepare the antibody against BmAIy/REF. Immuno-fluorescence examination showed that BmAly/REF was distributed in both cytoplasm and nucleus of the cell, but mainly located in the nucleus. Microarray data analysis showed that Bmaly/ref gene was highly expressed in tissues of the day-3 5th instar larvae of the silkworm. The results suggest that BmAly/ REF is likely to play important roles in RNA nuclear export, which lays a good foundation to further investigate the function of BmAly/REF.关键词
家蚕/BmAly/REF/基因克隆/序列分析/细胞定位/基因芯片Key words
Bombyx mori/ BmAly/REF/ gene cloning/ sequence analysis/ cellular localization/ microarray分类
生物科学引用本文复制引用
钟金凤,曹广力,薛仁宇,贡成良..家蚕Aly/REF的基因克隆、序列分析及其细胞定位[J].昆虫学报,2011,54(7):746-753,8.基金项目
国家自然科学基金项目(31072085) （31072085）
江苏省自然科学基金项目(BK2009117) （BK2009117）
苏州市自然科学基金项目(YJG0907) （YJG0907）
苏州大学重大应用研究培育项目(Q3134991) （Q3134991）
