Molecular Cloning and Differential Expression of lhcf4Gene from Gametophytes of Saccharina japonica

doi:10.7668/hbnxb.2011.06.009

Abstract

Abstract: lhcf4gene full length cDNA of Saccharina japonicawas cloned by using RT-PCR technique with primers designed based on Laminaria saccharinalhcf4full 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 isoelectric point of 5． 36． ORFs of lhcf4gene and its homologs were conserved，while their UTRs were diverse according to results of similarity analysis． The predicted 3D structure of lhcf4contained 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 lhcf4protein was determined by overlapping its 3D structure with Xray diffraction structure of pea LHC-II monomer at 2． 5 resolution． The Neighbor-joining phylogenetic tree constructed by MEGA4． 0 revealed three significantly different lineages from lhc genes: chromophytes，green alga and prochlorococcus，which might be have the same ancestor from cyanobacteria． Otherwise， it also revealed that lhcf4genes of S． japonicaand L． saccharina， fcpb gene of Macrocystis pyrifera were orthologs while lhcf4， lhcF₁and lhcF7 genes of L． saccharina were Paralogs． Real-time PCR results confirmed that lhcf4gene of S． japonicadifferentially expressed between in female and male gametophytes．

Key words: Saccharina japonica, Gametophytes, lhcf4, Gene cloning, Sequence analysis, Differential expression

CLC Number:

BI Yan-hui, ZHOU Zhi-gang. Molecular Cloning and Differential Expression of lhcf4Gene from Gametophytes of Saccharina japonica[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(6): 41-49. doi: 10.7668/hbnxb.2011.06.009.

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