PMID: 32696746Jul 23, 2020Paper

Prokaryotic expression of PE8 protein from Mycobacterium tuberculosis (H37Rv) and preparation of its polyclonal antibody in rabbits

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Tao XuHongtao Wang

Abstract

Objective To clone proline-glutamate 8 (PE8) gene segment from Mycobacterium tuberculosis (H37Rv), construct the recombinant plasmid pET28a-PE8, express recombinant PE8 protein, and prepare its polyclonal antibody. Methods Using a standard homologous recombination cloning technology, we cloned the PE8 gene into the prokaryotic vector pET28a. After sequence confirmation, it was transformed into E. coli BL21 (DE3) and treated with 0.5 mmol/L isopropyl-beta-D-thiogalactopyranoside (IPTG) to induce protein expression. We purified and renatured the recombinant PE8 protein, and immunized New Zealand rabbits to prepare the polyclonal antibody. Antibody titer was determined by indirect ELISA and the specificity was evaluated by Western blot analysis. Results The recombinant plasmid pET28a-PE8 was successfully constructed, and the PE8 protein was primarily expressed in an inclusion body in E. coli. After renaturation and purification, a purity of about 90% of the recombinant protein was achieved. The titer of the polyclonal antibody was higher than 1:430 080. The polyclonal antibody could specifically recognize the recombinant PE8 protein. Conclusion We have successfully expressed and purified recombinant PE8 protein, which can be furth...Continue Reading

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Shaoying GaoJunmin Luo
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Qingyuan ZhangPengfei Jiang
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Bingbing WangLifang Zhang
© 2021 Meta ULC. All rights reserved