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A preferable approach to clone hLIF cDNA from the genomic DNA Electron. J. Biotechnol.
Cui,Yihong; Zhu,Guangqin; Chen,Qiuju; Wang,Yunfei; Yang,Mingming; Song,Yuxuan; Wang,Jiangang; Cao,Binyun.
Complementary DNA (cDNA) is valuable for investigating protein structure and function in the research of life science, but it is difficult to obtain by traditional reverse transcription. In this study, we employed a novel strategy to clone the human leukemia inhibitory factor (hLIF) gene cDNA from genomic DNA directly isolated from the mucous membrane of mouth. The hLIF sequence can be acquired within a few hours by means of amplification of each exon and splicing using overlap-PCR. Thus, the new approach developed in this study is simple, time- and cost-effective, and it is not limited to particular gene expression levels of each tissue.
Tipo: Journal article Palavras-chave: CDNA cloning; HLIF; Oral mucous membrane; Overlap PCR.
Ano: 2011 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300012
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Molecular cloning and sequence analysis of growth hormone cDNA of Neotropical freshwater fish Pacu (Piaractus mesopotamicus) Genet. Mol. Biol.
Pinheiro,Janeth Silva; Wolff,José Luiz Caldas; Araújo,Ronaldo de Carvalho; Hilsdorf,Alexandre Wagner Silva.
RT-PCR was used for amplifying Piaractus mesopotamicus growth hormone (GH) cDNA obtained from mRNA extracted from pituitary cells. The amplified fragment was cloned and the complete cDNA sequence was determined. The cloned cDNA encompassed a sequence of 543 nucleotides that encoded a polypeptide of 178 amino acids corresponding to mature P. mesopotamicus GH. Comparison with other GH sequences showed a gap of 10 amino acids localized in the N terminus of the putative polypeptide of P. mesopotamicus. This same gap was also observed in other members of the family. Neighbor-joining tree analysis with GH sequences from fishes belonging to different taxonomic groups placed the P. mesopotamicus GH within the Otophysi group. To our knowledge, this is the first GH...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Piaractus mesopotamicus; Growth hormone; CDNA cloning; Sequencing.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000200037
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