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绵羊粒细胞-巨噬细胞集落刺激因子(GM-CSF)cDNA的克隆与测序

Cloning and sequencing of the cDNA for ovine granulocyte-macrophage colony-stimulating factor (GM-CSF).

作者信息

O'Brien P M, Rothel J S, Seow H F, Wood P R

机构信息

CSIRO Division of Animal Health, Animal Health Research Laboratory, Parkville, Victoria, Australia.

出版信息

Immunol Cell Biol. 1991 Feb;69 ( Pt 1):51-5. doi: 10.1038/icb.1991.8.

Abstract

Colony-stimulating factors (CSFs) are not only regulators of haemopoiesis but can also enhance the function of mature myeloid cells, and are therefore potential immune adjuvants. By use of the polymerase chain reaction (PCR) with primers based on the bovine granulocyte-macrophage CSF (GM-CSF) sequence, we have amplified the cDNA for ovine GM-CSF, produced from crude mRNA extracted from alveolar macrophages. The PCR product was cloned into pUC119, and electroporated into Escherichia coli. The complete nucleotide sequence of two clones, and the partial sequence of eight others, was determined. At the nucleotide and amino acid levels, the ovine and bovine GM-CSF sequences are 91% and 81% homologous, respectively.

摘要

集落刺激因子(CSFs)不仅是造血的调节因子,还能增强成熟髓样细胞的功能,因此是潜在的免疫佐剂。通过使用基于牛粒细胞-巨噬细胞集落刺激因子(GM-CSF)序列的引物进行聚合酶链反应(PCR),我们从肺泡巨噬细胞提取的粗制mRNA中扩增出了绵羊GM-CSF的cDNA。将PCR产物克隆到pUC119中,并电穿孔导入大肠杆菌。测定了两个克隆的完整核苷酸序列以及另外八个克隆的部分序列。在核苷酸和氨基酸水平上,绵羊和牛GM-CSF序列的同源性分别为91%和81%。

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