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2002•Zhongguo yufang shouyi xuebaoRequires access

Cloning and Sequencing of Porcine IL-10

Hailong Guo

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Abstract

The total RNA was extracted from pig peripheral blood mononuclear cells (PBMCs) stimulated by LPS.Porcine IL-10 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The gene was cloned into pUC18 vector and identified by PCR and digestion by BamHI and HindIII.Then one of the positive recombinant clones was sequenced and analyzed.The result showed that the gene has only one ORF,which is composed of 528 nucleosides.The predicted peptide included 175 amino acids.It also showed that the sequence was 84%,79% and 79% identity to that of human,rat and mouse,respectively.The sequence was just the same as the published one from GenBank.So the porcine IL-10 was cloned from porcine PBMCs successfully.

About this research paper

What this paper is about

The total RNA was extracted from pig peripheral blood mononuclear cells (PBMCs) stimulated by LPS.Porcine IL-10 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The gene was cloned into pUC18 vector and identified by PCR and digestion by BamHI and HindIII.Then one of the positive recombinant clones was sequenced and analyzed.The result showed that the gene has only one ORF,which is composed of 528 nucleosides.The predicted peptide included 175 amino acids.It also showed that the sequence was 84%,79% and 79% identity to that of human,rat and mouse,respectively.The sequence was just the same as the published one from GenBank.So the porcine IL-10 was cloned from porcine PBMCs successfully.

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Available abstract

The total RNA was extracted from pig peripheral blood mononuclear cells (PBMCs) stimulated by LPS.Porcine IL-10 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The gene was cloned into pUC18 vector and identified by PCR and digestion by BamHI and HindIII.Then one of the positive recombinant clones was sequenced and analyzed.The result showed that the gene has only one ORF,which is composed of 528 nucleosides.The predicted peptide included 175 amino acids.It also showed that the sequence was 84%,79% and 79% identity to that of human,rat and mouse,respectively.The sequence was just the same as the published one from GenBank.So the porcine IL-10 was cloned from porcine PBMCs successfully.

Key concepts: Biology, Molecular biology, GenBank, Cloning (programming), Gene, Recombinant DNA, Peripheral blood mononuclear cell, HindIII

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