pBR322复制动力学及其对生长速率的依赖性分析。

Analysis of pBR322 replication kinetics and its dependency on growth rate.

作者信息

Kim B G, Shuler M L

机构信息

Department of Food Science and Technology, Cornell University, Ithaca, New York 14853, USA.

出版信息

Biotechnol Bioeng. 1990 Jul;36(3):233-42. doi: 10.1002/bit.260360304.

DOI:10.1002/bit.260360304

PMID:18595073

Abstract

Accurate estimates of plasmid copy number in a cell are a prerequisite for predicting plasmid stability and protein production. A refined version of a structured model for the pBR322 plasmid replication mechanism is described. The model is capable of accurately predicting pBR322 plasmid copy number in Escherichia coli B/r for a wide range of growth rates. The refinements include better estimates of promoter strength, the degradation rate of RNA species, binding constant of RNAI-RNAII reaction, and dependency of promoter strength on growth rate. The predictions of the model are verified by recent experimental observations but differ from some previous reports. This model can also be used to predict the binding constant of the RNAI-RNAII reaction of ColE1 type plasmids. At 37 degrees C, the binding constant is estimated to be 77 +/- 11 x 10(-13) mL/molecule-h for pBR322.

摘要

准确估计细胞中的质粒拷贝数是预测质粒稳定性和蛋白质产量的前提条件。本文描述了pBR322质粒复制机制结构化模型的改进版本。该模型能够在很宽的生长速率范围内准确预测大肠杆菌B/r中pBR322质粒的拷贝数。改进之处包括对启动子强度、RNA种类的降解速率、RNAI-RNAII反应的结合常数以及启动子强度对生长速率的依赖性进行了更好的估计。该模型的预测结果得到了近期实验观察的验证，但与之前的一些报道有所不同。该模型还可用于预测ColE1型质粒RNAI-RNAII反应的结合常数。在37摄氏度时，pBR322的结合常数估计为77±11×10⁻¹³ mL/分子·小时。