利用三质粒载体系统对有害蛋白的可控表达进行建模。

Modeling of the controlled expression of a harmful protein by a three-plasmid harboring system.

作者信息

Unzaga T V, Díaz-Ricci J C, Rhee J I, Hernández M R, Schügerl K

机构信息

Instituto de Ingeniería Química, Facultad de Ciencias Exactas y Tecnología, Avda. Independencia 1800, 4000 Tucumán, Argentina.

出版信息

Biotechnol Bioeng. 2002 Dec 5;80(5):544-51. doi: 10.1002/bit.10412.

DOI:10.1002/bit.10412

PMID:12355465

Abstract

A genetically structured mathematical model was developed and used to evaluate the influence of molecular parameters involved in the expression of a harmful recombinant protein (SPA::EcoRI). The system consists of the controlled expression of the endonuclease EcoRI cloned in the plasmid pMTC48. The control is exerted by the lambda CI repressor expressed from the plasmid pRK248cIts. The deleterious effect of the activity of the enzyme EcoRI on the host DNA is prevented by the action of the EcoRI methylase that is expressed constitutively from a third plasmid, pEcoR4. The model includes molecular mechanisms involved in the regulation of the expression of these genes and is used to determine cultural conditions that maximize the production of the recombinant protein.

摘要

构建了一个基因结构数学模型，并用于评估参与有害重组蛋白（SPA::EcoRI）表达的分子参数的影响。该系统由克隆在质粒pMTC48中的内切核酸酶EcoRI的受控表达组成。控制由从质粒pRK248cIts表达的λCI阻遏物施加。通过从第三个质粒pEcoR4组成型表达的EcoRI甲基化酶的作用，可防止EcoRI酶活性对宿主DNA的有害影响。该模型包括参与这些基因表达调控的分子机制，并用于确定使重组蛋白产量最大化的培养条件。