Cloning and Identification of lhcf6 Gene Promoters from Saccharina japonica Gametophytes

doi:10.7668/hbnxb.2014.01.012

Abstract

Abstract: lhcf6 gene 5'-flanking sequences were isolated respectively from genomic DNA of female and male gametophytes of Saccharina japonica by nested PCR using genomic walking method. The length of the fragments were 932 bp in female and 901 bp in male. Sequence alignment showed a low similarity of 33. 3% between the two fragments,indicating this gene had different upstream regulatory region in female and male gametophytes. Cis-elements prediction by web server PlantCARE and Softberry showed both of them contained TATA box,circadian responsive element MBS,as well as light responsive elements MNF1 and GAG-motif. Otherwise,some elements were specific to female or male lhcf6,such as LHC gene promoter typical element AT-rich sequence was specific to female and circadian element circadian was specific to male. These elements might take part in the regulation of lhcf6 expression by light quality,light intensity and circadian. These two 5'-flanking sequences were fused with the reporter gene GFP to construct plant expression vectors,named pCAMBIA1304-Flhcf6 and pCAMBIA1304-Mlhcf6. The recombinant vectors as well as vector pCAMBIA1304 as positive control were transferred to the green alga Chlamydomonas reinhardtii by electroporation. GFP was observed under laser confocal fluorescence microscopy. Fluorescence signal was detected in cells containing pCAMBIA1304-Flhcf6 and pCAMBIA1304-Mlhcf6 indicating both the two cloned 5'-flanking sequences showed promoter activity.

Key words: Saccharana japonaca, Gametophytes, Lhcf6 gene, Promoter, Green fluorescent protein

CLC Number:

NING Pu, BI Yan-hui, ZHOU Zhi-gang. Cloning and Identification of lhcf6 Gene Promoters from Saccharina japonica Gametophytes[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(1): 62-68. doi: 10.7668/hbnxb.2014.01.012.

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