毛白杨木质素合成酶基因F5H克隆与生物信息学分析

克隆毛白杨阿魏酸-5-羟基化酶(F5H)基因全长序列,并通过生物信息学分析,为进一步研究F5H基因功能提供基础.利用RT-PCR方法成功从毛白杨741组培苗叶片中克隆了F5H基因,并对F5H基因的核苷酸序列及推导的氨基酸序列进行分析,包括疏水性-亲水性、氨基酸翻译后修饰、亚细胞定位、跨膜结构域、蛋白质二级三级功能结构域等进行分析预测和推断.获得的基因长度为l 686 bp,编码区1542 bp,与已公布的毛果杨F5H mRNA序列(GenBank登录号为AJ010324)的编码区相似性达97.45％,并与其他植物的该基因序列具有同源性.F5H是亲水性较强的蛋白,整条肽链有1个跨膜结构域;通过亚细胞定位表明F5H主要位于质体中,并含有磷酸化位点有19个.F5H含有丰富的二级结构,卷曲占44.44％、折叠占7.02％、螺旋占48.54％,并预测了其三维结构.分析结果显示,植物的阿魏酸-5-羟基化酶是一个具有跨膜结构域的亲水性蛋白,在内质网等分泌途径中表达.

Cloning and bioinformatics analysis on Lignin synthetase gene Ferulate-5-hydroxylase of Populus tomentosa

It provides a basis for further study of the gene function of Ferulate-5-hydroxylase(F5H) to clone and make bioinformatics analysis of F5H gene in Populus tomentosa. F5H gene was cloned from P. tomentosa leaves by RT-PCR method. The physical chemistry characteristics including hydrophobsity, post -translational modification, subcellular localization, transmembrane peptide, secondary and 3D structure of F5H were analyzed by applications on-line. The length of acquired sequence was 1 686 bp and the sequence in coding region is 1542bp. Compared with the coding region of F5H mRNA in P. trichocarpa (Accession Number: AJ010324), the resemblance was 97.45% . The results of subcellular localization showed that the F5H mainly lies in plasmid, which was analyzed by prediction tools. The secondary structure of F5H was various: 44.44% of Alpha helix, 7.02% of Extended strand and 48.54% of Random coil. 3D structure of F5H was predicted by SWISS-MODE. The results show that F5H is a hydrophilic protein with two transmembrane peptides and expresses in the endoplasmic reticulum and other secretory pathways.