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- W2364155627 abstract "The β-agarase gene,agaD02,was amplified by PCR using genomic DNA of Agarivorans albus strain QM38 as a template.The agaD02 gene consists of an open reading frame of 2868 bp encoding β-agarase,a protein of 955 amino acids and a molecular weight of 106 kD.The PI of the agarase is 4.87.The accession numbers for agaD02 in GenBank was EF199908.We found that the agaD02 gene shared 98.7% sequence identity to the agaB gene of Vibrio sp.JT0107 and 97.4% identity to the agaE gene of Vibrio sp.PO-303.The deduced amino acid sequence of the agaD02 gene was compared with entries in the DDBJ database.According to the sequence of agaD02 gene,a pair of primers were designed and synthesized.After PCR amplification,the product was cloned into pMD19-T simple vector using TA clon-ing.The recombinants were sequenced and identified by restrictive endonuclease digestion.The target gene sequences were then subcloned into a highly efficient eukaryotic expression vector pET24a(+).After confirmation by double restric-tive endonuclease digestion,the expression vectors were transformed into E.coli BL21(DE3).The recombinants E.coli BL21(pET24a-agaD02) were induced by IPTG on Petri dish and were stained with Lugol iodide solution after being cultured for 24 h at 37℃.A clear zone was observed around the colony of the recombinants E.coli,confirming that the pET24a-agaD02 expression vectors were successfully constructed." @default.
- W2364155627 created "2016-06-24" @default.
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- W2364155627 date "2010-01-01" @default.
- W2364155627 modified "2023-09-28" @default.
- W2364155627 title "Cloning and expression of the beta-agarase gene agaD02 from Agarivorans albus QM38" @default.
- W2364155627 hasPublicationYear "2010" @default.
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