Molecular cloning and characterization of the allophycocyanin gene from Spirulina platensis C1

การโคลนยีนแอลโลไฟโคไซยานินจากสาหร่ายสีน้ำเงินแกมเขียว SPIRULINA PLATENSIS C1

Name: Dauenpen Meesapyodsuk

Organization : King Mongkut's University of Technology Thonburi

keyword: Spirulina platensis Ci

; allophycocyanin gene

; apcABC operon

; สาหร่ายสีน้ำเงินแกมเขียว

; Spirulina

Abstract: The apcABC genes encoding a, p subunits of allophycocyaninand partial part of core linker protein were isolated from S. platensis Cl.Based on sequence comparisons of apt genes from SynechococcusPCC7002, Synechococcus PCC6301 and Cyanophora paradoxa, twohighly conserved regions were selected for the synthesis of 2oligonucleotides, primers for amplifying apt gene probe from genomicDNA of S. platensis Ci by polymerase chain reaction technique. Theamplified probe was used for screening S. platensis Ci genomic DNAlibrary. By homologous hybridization the upstream sequence of apcAand partial sequence of the coding region of apcA gene was obtained. Inorder to clone the entire apt genes, primer down in the apcC wasdesigned from the conserved sequence of the corresponding genes fromfive cyanobacteria (Synechococcus PCC6301, Synechocystis PCC6803,Synechococcus elongatus, FremyeZZa diplosiphon and Synechococcus’vPCC7002) and primer up from a part of the upstream sequence of apcAisolated from genomic DNA library. By using these two primers, anamplified DNA fragment of 1.8 kb was obtained, and its nucleotidesequence was determined and analysed. Two complete open readingframes of-qcA and-+zB (16 1 amino acids-for -each gene), and a partialone of apcC (36 amino acids) were found. The identity of the deducedamino acid sequence of the products of apcA and apcB to those ofSynechococcus elongatus, Cyanophora paradoxa and SynechocystisPCC6714 were 85,82 and 80% for apcA, and 93, 84 and 88% for apcB,respectively. For apcC, the identity of amino acid sequences between S.platensis Ci and Synechococcus elongatus, Synechocystis PCC67 14 andFremyella diplosiphon were 92, 92 and 92%, respectively. Expectedchromophore attachment sites were found at position 81 in both aminoacid sequences of apcA and apcB. The sequence analysis alsodemonstrated that the obtained genes form an operon, apcABC, with asingle transcription start site and one possible termination sitedownstream of apcB. In addition, by sequence comparison of theupstream region of apcA of S. platensis Ci with those from othercyanobacteria, it was found that the apcABC promoters lied in theconserved sequences centered at -50 and -10 with respect to the start oftranscription.These results suggest that the obtained genes are putativeapcABC operon of S. platensis Cl.

King Mongkut's University of Technology Thonburi

Address: กรุงเทพมหานคร (Bangkok)

Email: info.lib@mail.kmutt.ac.th

Name: Dr. Supaporn Cheevdhanarak

Role: Supervisor

Created: 1996

Issued: 2005-04-09

Modified: 2010-07-10

วิทยานิพนธ์/Thesis

ISBN: 9746219049

CallNumber: BIT174

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biotechnology

Grantor: King Mongkut's University of Technology Thonburi

©copyrights King Mongkut's University of Technology Thonburi

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