摘要
旨在建立麻疯树BIBAC文库筛选目的基因快捷高效的方法,并对其克隆的叶绿体ω-3脂肪酸去饱和酶基因进行生物信息学分析。本研究利用高密度杂交膜从麻疯树BIBAC文库中筛选到含有叶绿体ω-3脂肪酸去饱和酶基因的克隆,对其测序并用生物信息软件对获得全长序列及编码的蛋白质序列的特性进行分析。将筛选到的阳性克隆进行测序,并将该序列与麻疯树叶绿体ω-3脂肪酸去饱和酶基因进行比对,相似性达到99%,证实此克隆为含叶绿体ω-3脂肪酸去饱和酶基因(JcFAD7)的BIBAC克隆。麻疯树叶绿体ω-3脂肪酸去饱和酶基因长度为2655bp,含有8个外显子和7个内含子,编码525个氨基酸。该基因编码的蛋白质为不具有信号肽的亲水性蛋白,二级结构元件多为α-螺旋和无规则卷曲,含有2个大的跨膜区域,在麻疯树中为单拷贝。本研究建立了高密度杂交膜从BIBAC文库中筛选目的基因快速高效的方法,同时也验证了该方法的可行性并对克隆到的基因进行了生物信息学分析。
The goal of this research is to establish a fast way to identify positive clones from Jatropha BIBAC library and bioinformatics analysis of chloroplast ω-3 fatty acid desaturase.Screening of a BIBAC library of Jatropha curcas L.using high-density clone filters with overgo designed from chloroplast ω-3 fatty acid desaturase,one positive clone was found and sequenced,the sequence of nucleic acid and protein were analyzed by bioinformatics software,respectively.The sequencing result suggested the nucleotide sequences of this positive clone showed 99% identity to chloroplast ω-3 fatty acid desaturase（JcFAD7）,which confirmed the fact that this clone contains gene chloroplast ω-3 fatty acid desaturase.Jatropha chloroplast ω-3 fatty acid desaturase was 2655 bp,consisting of 8 exons and 7 introns,and could be translated into 525 amino acids.The bioinformatics analysis result showed that JcFAD7 was hydrophilic with no leader peptides,α-helixs and random coils were dominant elements in secondary structure and it contained two trans-membrance sections.This research not only confirmed the utility of finding a gene by screening a large-insert genome library,but also found a way to locate interested gene in Jatropha BIBAC library and bioinformatics analysis of chloroplastω-3 fatty acid desaturase.
出处
《中国农学通报》
CSCD
2012年第31期21-28,共8页
Chinese Agricultural Science Bulletin
基金
国际合作项目"燃料作物共同研究
种质收集与繁殖"(2008KR0395)
