华北农学报2011，Vol.26Issue(6)：41-49,9.
海带lhcf4基因克隆和配子体差异表达研究
Molecular Cloning and Differential Expression of lhcf4 Gene from Gametophytes of Saccharina japonica
摘要
Abstract
Ihcf4 gene full length cDNA of Saccharina japonica was cloned by using RT-PCR technique with primers designed based on Laminaria saccharina Ihcf4 full length cDNA sequence ( accession number: AF226860 ). Its ORF encoded a precursor protein containing 218 amino acids with a molecular weight of 23. 11 kDa and an isoe-lectric point of 5. 36. ORFs of Ihcf4 gene and its homologs were conserved, while their UTRs were diverse according to results of similarity analysis. The predicted 3D structure of LHCF4 contained 4 α-helixes in which the first and third helix were transmembrane helices, the second one was a little shorter and embedded in the thylakoid membrane, the fourth one was a short helix on the lumenal side. Chlorophyll a binding sites and amina acid residues forming salt-bridge between the first and third helix were conserved in chromophytes, green algae and higher plants. The spatial distribution of chlorophyll a in LHCF4 protein was determined by overlapping its 3D structure with X-ray diffraction structure of pea LHC-Ⅱ monomer at 2. 5(A) resolution. The Neighbor-joining phylogenetic tree constructed by MEGA4.0 revealed three significantly different lineages from Ihc genes; chromophytes, green alga and prochlorococcus, which might be have the same ancestor from cyanobacteria. Otherwise, it also revealed that Ihcf4 genes of S. Japonica and L. Saccharina ,fcpb gene of Macrocystis pyrifera were orthologs while Ihcf4 , Ihcf1 and lh'cF7 genes of L. Saccharina were Paralogs. Real-time PCR results confirmed that Ihcf4 gene of 5. Japonica differentially expressed between in female and male gametophytes.关键词
海带/配子体/lhcf4/基因克隆/序列分析/差异表达Key words
Saccharina japonica / Gametophytes/ Ihcf4 / Gene cloning/ Sequence analysis/ Differential expression分类
生物科学引用本文复制引用
毕燕会,周志刚..海带lhcf4基因克隆和配子体差异表达研究[J].华北农学报,2011,26(6):41-49,9.基金项目
上海高校选拔培养优秀青年教师科研专项基金(SSC09017) （SSC09017）
上海海洋大学博士科研启动基金(B-8212-09-000504) （B-8212-09-000504）
上海市自然科学基金(KW-201024727) （KW-201024727）
