Isolation and DNA sequence analysis of distantly related fatty acid and retionol binding protein gene of Setaria digitata

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dc.contributor.authorYATAWARA, MDMLDKen_US
dc.date.accessioned2011-02-14T15:16:25Z
dc.date.available2011-02-14T15:16:25Z
dc.date.created2003en_US
dc.date.issued2003en_US
dc.descriptionDissertation: M.Sc., University of Colombo: UC(MED), 2003.en_US
dc.description.abstractPreviously sequenced cDNA clone (designated PSD38) of Seteria digitata revealed a 102 bp region that had high homology (76 percent) to Fatty Acid and Retinol binding protein (FAR) gene of Brugia malayi. Rapid Amplification of 5' cDN Ends (RACE) method was performed on mRNA of Seteria digitata using primers derived from the above sequence. To obtain the promoter region of that gene, Genome Walker DNA Walking method was used. cDNA synthesized from Seteria digitata mRNA gave a 700 base pair fragment designated race-1. This fragment (race-1) was completely sequenced by PCR cycle sequencing. The sequence showed (64 percent) AT richness. When this sequence was aligned with NCBI GeneBank data a region of 109 bp showed a high degree (100 percent) of homology to Brugia malayi FAR protein gene sequence. Amino acid sequence deduced from this region (36 amino acids) had 100 percent similarity with the Brugia malayi FAR protein sequence. Single open reading frame of the race-1 sequence could not be obtained due to the presence of stop codons. The contig (cl) constructed from the race-1 sequence and the PSD38 sequence showed two regions that had high homology to Brugia nalayi FAR protein gene. A 109 bp (towards the 5' end) and a 102 bp (towards the 3' end) respectively. Seteria digitata genome walker library was constructed by cleaving genomic DNA with restriction enzymes. PCR amplification of this library using gene specific primers revealed a 636 bp region in the Dra 1 library. This fragment was cloned in pGEM T easy vector system 11 and recombinant clone designated DL-1 GW was sequenced completely. The sequence showed 74 percent AT richness displaying the characteristic observed in nematode genomes. The cloned sequenced was aligned with NCBI GenBank data showed a region of 100bp having 84 percent homology with B.malayi FAR protein gene sequence. The sequence contained ATG initiation codon and putative TATAA box indication the promoter region in the 5' end. When the DL-1 GW sequence was aligned with race-1 sequence a 100 bp region in the center and a 37bp region at the 3' end showed 100 percent homology. The resultindicate that the characterized sequences of Seteria digitata showed regions of sililary to the Brugia malayi FAR protein gene. The amino acids deduced from the homologous regions constitute 40 percent of the Brugia malayi FAR protein.en_US
dc.identifier.citationYATAWARA, MDMLDK, Isolation and DNA sequence analysis of distantly related fatty acid and retionol binding protein gene of Setaria digitata, University of Colombo UC(MED), 2003: 92p.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/129486
dc.language.isoen_USen_US
dc.publisherUniversity of Colombo: UC(MED).en_US
dc.rightsUniversity of Colombo, UC(MED): Sri Lanka HELLIS Networken_US
dc.source.urihttps://hellis.srilanka.healthrepository.orgen_US
dc.subjectFilarioideaen_US
dc.subject.meshSequence Analysis, DNAen_US
dc.subject.meshRetinol-Binding Proteinsen_US
dc.subject.meshSequence Analysis, DNA-methodsen_US
dc.subject.meshDNA, Helminth-analysisen_US
dc.subject.meshSetaria Nematode-isolation \& purificationen_US
dc.subject.meshSetaria Nematodeen_US
dc.subject.meshSetariasis-parasitologyen_US
dc.subject.meshSetaria Nematode-geneticsen_US
dc.titleIsolation and DNA sequence analysis of distantly related fatty acid and retionol binding protein gene of Setaria digitataen_US
dc.typeThesisen_US
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