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利用针对在大肠杆菌中产生的融合蛋白制备的抗血清对逆转录病毒pol基因产物进行分析。

Analysis of retroviral pol gene products with antisera raised against fusion proteins produced in Escherichia coli.

作者信息

Tanese N, Roth M J, Goff S P

出版信息

J Virol. 1986 Aug;59(2):328-40. doi: 10.1128/JVI.59.2.328-340.1986.

Abstract

Portions of the pol gene of Moloney murine leukemia virus (MuLV) were expressed as fusion proteins in Escherichia coli, and the purified proteins were used to elicit antibodies in Escherichia coli, and the purified proteins were used to elicit antibodies in rabbits. The sera were used to examine the mature pol gene products contained in virion particles and identified the reverse transcriptase and a second protein, P46pol, encoded by the 3' portion of the gene. The P46 protein was not phosphorylated and was present at the same molar abundance as the reverse transcriptase. The sera were also used to detect the Pr200gag-pol intracellular precursor protein and to analyze its processing to the mature forms. The proteins formed by several Moloney MuLV mutants were analyzed. Further tests revealed cross-reactivity with Friend MuLV and feline leukemia virus proteins, but not with avian retrovirus proteins.

摘要

莫洛尼鼠白血病病毒(MuLV)的部分pol基因在大肠杆菌中表达为融合蛋白,纯化后的蛋白用于在兔体内诱导产生抗体。这些血清用于检测病毒粒子中所含的成熟pol基因产物,并鉴定了由该基因3'部分编码的逆转录酶和第二种蛋白P46pol。P46蛋白未被磷酸化,其摩尔丰度与逆转录酶相同。这些血清还用于检测Pr200gag-pol细胞内前体蛋白,并分析其加工成成熟形式的过程。对几种莫洛尼MuLV突变体形成的蛋白进行了分析。进一步的检测显示,这些血清与弗瑞德MuLV和猫白血病病毒蛋白有交叉反应,但与禽逆转录病毒蛋白无交叉反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2673/253082/dcc437abc3f8/jvirol00107-0147-a.jpg

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