Tanese N, Prasad V R, Goff S P
Department of Biochemistry and Molecular Biophysics, Columbia University, College of Physicians and Surgeons, New York, NY 10032.
DNA. 1988 Jul-Aug;7(6):407-16. doi: 10.1089/dna.1.1988.7.407.
A collection of variant plasmids that express the human immunodeficiency virus (HIV) reverse transcriptase as trpE fusion proteins were generated and scored for their ability to produce stable, active proteins. Trimming portions of the viral pol gene resulted in dramatic increases in yield over earlier constructs; the accumulation of high levels of enzymatically active protein in this system was increased by the retention of the trpE sequences at the amino terminus. A new in situ gel activity assay was used to demonstrate that the major induced protein, containing approximately 68 kD of viral sequences, was the active species.
构建了一组表达人免疫缺陷病毒（HIV）逆转录酶作为trpE融合蛋白的变异质粒，并对其产生稳定活性蛋白的能力进行了评分。对病毒pol基因的部分进行修剪，使得产量比早期构建体有显著提高；通过在氨基末端保留trpE序列，该系统中高水平酶活性蛋白的积累增加。一种新的原位凝胶活性测定法被用于证明主要的诱导蛋白（包含约68kD的病毒序列）是活性物种。
