Expression of Leptospira membrane proteins Signal Peptidase (SP) and Leptospira Endostatin like A (Len A) in BL-21(DE3) is toxic to the host cells

IF 3.5 Q3 Biochemistry, Genetics and Molecular Biology
Padikara K. Satheeshkumar , Prasannan V. Anu , Mohmed I. Junaida , Madathiparambil G. Madanan , Tennison Jebasingh , Ananthakrishnan J. Nair , Gangaprasad A. Nair , Govinda Pillai M. Nair , Perumana R. Sudhakaran
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引用次数: 2

Abstract

Heterologous expression of Integral Membrane Proteins (IMPs) is reported to be toxic to the host system in many studies. Even though there are reports on various concerns like transformation efficiency, growth properties, protein toxicity, inefficient expression and protein degradation in IMP overexpression, no studies so far addressed these issues in a comprehensive way. In the present study, two transmembrane proteins of the pathogen Leptospira interrogans, namely Signal peptidase (SP), and Leptospira Endostatin like A (Len-A) were taken along with a cytosolic protein Hydrolase (HYD) to assess the differences in transformation efficiency, protein toxicity, and protein stability when over expressed in Escherichia coli (E. coli). Bioinformatics analysis to predict the transmembrane localization indicated that both SP and Len are targeted to the membrane. The three proteins were expressed in full length in the E. coli expression strain, BL 21 (DE3). Significant changes were observed for the strains transformed with IMP genes under the parameters analysed such as, the transformation efficiency, survival of colonies on IPTG-plate, culture growth kinetics and protein expression compared to the strain harbouring the cytosolic protein gene.

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钩端螺旋体膜蛋白信号肽酶(SP)和钩端螺旋体内皮抑素样A (Len A)在BL-21(DE3)中的表达对宿主细胞具有毒性
在许多研究中,整体膜蛋白(IMPs)的异源表达被报道对宿主系统有毒性。尽管IMP过表达的转化效率、生长特性、蛋白质毒性、低效表达和蛋白质降解等问题都有报道,但目前还没有研究全面解决这些问题。本研究以致病菌钩端螺旋体(Leptospira疑问)的两种跨膜蛋白信号肽酶(SP)和内皮抑素样A (Len-A)与细胞质蛋白水解酶(HYD)结合,评估了在大肠杆菌(E. coli)中过表达时转化效率、蛋白毒性和蛋白稳定性的差异。预测跨膜定位的生物信息学分析表明SP和Len都是靶向细胞膜的。这3种蛋白在大肠杆菌表达菌株bl21 (DE3)中完整表达。转染IMP基因的菌株在转化效率、菌落在iptg板上的存活率、培养生长动力学和蛋白表达等参数上与转染细胞质蛋白基因的菌株相比有显著变化。
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来源期刊
Journal of Genetic Engineering and Biotechnology
Journal of Genetic Engineering and Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
5.70
自引率
5.70%
发文量
159
审稿时长
16 weeks
期刊介绍: Journal of genetic engineering and biotechnology is devoted to rapid publication of full-length research papers that leads to significant contribution in advancing knowledge in genetic engineering and biotechnology and provide novel perspectives in this research area. JGEB includes all major themes related to genetic engineering and recombinant DNA. The area of interest of JGEB includes but not restricted to: •Plant genetics •Animal genetics •Bacterial enzymes •Agricultural Biotechnology, •Biochemistry, •Biophysics, •Bioinformatics, •Environmental Biotechnology, •Industrial Biotechnology, •Microbial biotechnology, •Medical Biotechnology, •Bioenergy, Biosafety, •Biosecurity, •Bioethics, •GMOS, •Genomic, •Proteomic JGEB accepts
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