油菜蔗糖转化酶基因的电子克隆和生物信息学分析

运用电子克隆技术获得油菜中一个蔗糖转化酶基因eDNA序列，同时根据此段序列设计引物以油菜eDNA为模板进行扩增。经测序得到证实。采用生物信息学方法，对该基因编码蛋白从氨基酸组成、基本理化性质、跨膜结构域、信号肽导肽、疏水性／亲水性、二级结构、亚细胞定位等方面进行了预测和分析。结果表明：该基因eDNA序列长度为2150bp，包含一个1779bp开放阅读框，编码592个氨基酸；该编码蛋白含有蔗糖转化酶的多个典型的保守结构域。同源比对分析显示，该基因编码的氨基酸序列与拟南芥等植物的蔗糖转化酶基因具有高度的相似性，进一步确定该蛋白为蔗糖蛋白酶。研究结果为该基因进一步的实验克隆，表达分析，功能鉴定奠定基础。

In silico cloning and bioinformatics analysis of invertase gene in Brassica napus

A Invertase gene was obtained from Brassica napus by using the in silico cloning technique. At the same time we amplified this sequence with rape cDNA as a template using primers designed according to the sequence of cDNA. Sequencing confirmed the reliability of the results. The protein of this gene was also predicted and studied through the bioinformatics analysis method, including the composition of amino acid sequence, basic physical and chemical properties, transmembrane region, signal peptide, guide peptide, hydrophobicity and hydrophilicity, sec- ondary structure of protein, subcellular localization and so on. The results showed that the cDNA length of the gene was 2150bp, which included a 1 779bp open reading frame（ORF） and encoded 592 amino acids residue. The en- coding protein of the INV gene possessed several typical conserved domains. Homology comparison and phylogenet- ic analysis showed that the amino acid encoded by INV gene in Brassica napus was highly homologous with those encoded by INV gene in Arabidopsis thaliana and so on. The above results determine the protein is invertase and these will provide the basis for molecular cloning , expression analysis and function identification of INV genes in Brassica napus further.