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2014•Progress in Veterinary MedicineRequires access

Cloning,Sequencing and Eukaryotic Expression of Bovine Interleukin-10 Gene in CHO Cells

Wen De-ba

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Abstract

Bovine IL-10(bIL-10)gene was amplified by RT-PCR from peripheral blood lymphocytes of healthy bovine in this study.The sequencing results indicated that the bovine IL-10gene was 537bp in full length,and translated by 179amino acids,with a very stable genetic conservation.This gene was inserted into the eukaryotic expression vector pcDNA3/V5-His Vectors A,and successfully constructed the eukaryotic expression vector of pcDNA/V5-bIL-10,and then transfected into the CHO cells for expression. Then expression products in culture of supernatant and precipitate were identified by SDS-PAGE and Western blot methods,which had a molecular weight of approximately 20ku.These experiments lay a foundation for further study on the biological characteristics and gene polymorphism of the bIL-10.

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What this paper is about

Bovine IL-10(bIL-10)gene was amplified by RT-PCR from peripheral blood lymphocytes of healthy bovine in this study.The sequencing results indicated that the bovine IL-10gene was 537bp in full length,and translated by 179amino acids,with a very stable genetic conservation.This gene was inserted into the eukaryotic expression vector pcDNA3/V5-His Vectors A,and successfully constructed the eukaryotic expression vector of pcDNA/V5-bIL-10,and then transfected into the CHO cells for expression. Then expression products in culture of supernatant and precipitate were identified by SDS-PAGE and Western blot methods,which had a molecular weight of approximately 20ku.These experiments lay a foundation for further study on the biological characteristics and gene polymorphism of the bIL-10.

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Available abstract

Bovine IL-10(bIL-10)gene was amplified by RT-PCR from peripheral blood lymphocytes of healthy bovine in this study.The sequencing results indicated that the bovine IL-10gene was 537bp in full length,and translated by 179amino acids,with a very stable genetic conservation.This gene was inserted into the eukaryotic expression vector pcDNA3/V5-His Vectors A,and successfully constructed the eukaryotic expression vector of pcDNA/V5-bIL-10,and then transfected into the CHO cells for expression. Then expression products in culture of supernatant and precipitate were identified by SDS-PAGE and Western blot methods,which had a molecular weight of approximately 20ku.These experiments lay a foundation for further study on the biological characteristics and gene polymorphism of the bIL-10.

Key concepts: Transfection, Molecular biology, Cloning (programming), Gene, Biology, Vector (molecular biology), Molecular cloning, Western blot

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Cloning,Sequencing and Eukaryotic Expression of Bovine Interleukin-10 Gene in CHO Cells — Research Paper | ScholarLens