Cloning and expression analysis of the chloroplast fructose- 1,6-bisphosphatase gene from Pyropia haitanensis
doi: 10.1007/s13131-014-0455-0
Cloning and expression analysis of the chloroplast fructose- 1,6-bisphosphatase gene from Pyropia haitanensis
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摘要: Fructose-1,6-bisphosphatase (FBPase) is one of the key enzymes in Calvin circle and starch biosynthesis. In this study, the full-length of cpFBPase gene from Pyropia haitanensis was cloned by using rapid amplification of cDNA ends (RACE) technology. The nucleotide sequence of PhcpFBPase consists of 1 400 bp, including a 5' untranslated region (UTR) of 92 bp, a 3' UTR of 69 bp, and an open reading frame (ORF) of 1 236 bp, which can be translated into a 412-amino-acid putative peptides with a molecular weight of 44.3 kDa and a theoretical pI of 5.23. Multiple sequence alignment indicated that the protein belonged to the chloroplast FBPase enzyme. Phylogenetic analysis showed that the protein assembled with the cpFBPase of a thermal tolerant unicellular red micro-algae Galdieria sulphuraria. Expression patterns analyzed by qRT-PCR revealed that the expression of PhcpFBPase gene in the thallus phage was 7-fold higher than in the conchocelis phage, which suggested the different mechanisms of inorganic carbon utilization among the different life phages of P. haitanensis. And the different response modes of PhcpFBPase mRNA levels to high temperature and desiccation stress indicated that PhcpFBPase played an important role in responsing to abiotic stress.
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关键词:
- Pyropiahaitanensis /
- fructose-1,6-bisphosphatase /
- genecloning /
- qRT-PCR /
- RACE
Abstract: Fructose-1,6-bisphosphatase (FBPase) is one of the key enzymes in Calvin circle and starch biosynthesis. In this study, the full-length of cpFBPase gene from Pyropia haitanensis was cloned by using rapid amplification of cDNA ends (RACE) technology. The nucleotide sequence of PhcpFBPase consists of 1 400 bp, including a 5' untranslated region (UTR) of 92 bp, a 3' UTR of 69 bp, and an open reading frame (ORF) of 1 236 bp, which can be translated into a 412-amino-acid putative peptides with a molecular weight of 44.3 kDa and a theoretical pI of 5.23. Multiple sequence alignment indicated that the protein belonged to the chloroplast FBPase enzyme. Phylogenetic analysis showed that the protein assembled with the cpFBPase of a thermal tolerant unicellular red micro-algae Galdieria sulphuraria. Expression patterns analyzed by qRT-PCR revealed that the expression of PhcpFBPase gene in the thallus phage was 7-fold higher than in the conchocelis phage, which suggested the different mechanisms of inorganic carbon utilization among the different life phages of P. haitanensis. And the different response modes of PhcpFBPase mRNA levels to high temperature and desiccation stress indicated that PhcpFBPase played an important role in responsing to abiotic stress.-
Key words:
- Pyropia haitanensis /
- fructose-1,6-bisphosphatase /
- gene cloning /
- qRT-PCR /
- RACE
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