Expression Vectors for the Rapid Purification of Recombinant Proteins in Bacillus subtilis

2007 ◽  
Vol 55 (2) ◽  
pp. 89-93 ◽  
Author(s):  
Hoang Duc Nguyen ◽  
Trang Thi Phuong Phan ◽  
Wolfgang Schumann
Keyword(s):  
Bacillus Subtilis ◽  
Recombinant Proteins ◽  
Expression Vectors ◽  
Rapid Purification

scholarly journals Integrative expression vectors with Pgrac promoters for inducer-free overproduction of recombinant proteins in Bacillus subtilis

2020 ◽  
Vol 28 ◽  
pp. e00540
Author(s):  
Dinh Thi Minh Tran ◽  
Trang Thi Phuong Phan ◽  
Thanh Thi Ngoc Doan ◽  
Thuoc Linh Tran ◽  
Wolfgang Schumann ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Recombinant Proteins ◽  
Expression Vectors

State of the art in the production of transgenic goats

2004 ◽  
Vol 16 (4) ◽  
pp. 465 ◽  
Author(s):  
H. Baldassarre ◽  
B. Wang ◽  
C. L. Keefer ◽  
A. Lazaris ◽  
C. N. Karatzas
Keyword(s):  
Recombinant Proteins ◽  
Traditional Method ◽  
Nuclear Transfer ◽  
State Of The Art ◽  
Expression Vectors ◽  
Marketing Approval ◽  
Transfected Cells ◽  
Pronuclear Microinjection ◽  
Transgenic Goats

This review summarises recent advances in the field of transgenic goats for the purpose of producing recombinant proteins in their milk. Production of transgenic goats via pronuclear microinjection of DNA expression vectors has been the traditional method, but this results in low efficiencies. Somatic cell nuclear transfer has dramatically improved efficiencies in rates of transgenesis. Characterisation of transfected cells in vitro before use in nuclear transfer guarantees that kids born are transgenic and of predetermined gender. Using these platform technologies, several recombinant proteins of commercial interest have been produced, although none of them has yet gained marketing approval. Before these technologies are implemented in goat improvement programmes, efficiencies must be improved, costs reduced, and regulatory approval obtained for the marketing of food products derived from such animals.

scholarly journals Comparison of Expression Vectors for Transient Expression of Recombinant Proteins in Plants

2013 ◽  
Vol 31 (6) ◽  
pp. 1529-1538 ◽  
Author(s):  
Kausar Hussain Shah ◽  
Bachar Almaghrabi ◽  
Holger Bohlmann
Keyword(s):  
Transient Expression ◽  
Recombinant Proteins ◽  
Expression Vectors

scholarly journals Different expression levels of two KgmB-His fusion proteins

2005 ◽  
Vol 70 (12) ◽  
pp. 1401-1407 ◽  
Author(s):  
Sandra Markovic ◽  
Sandra Vojnovic ◽  
Milija Jovanovic ◽  
Branka Vasiljevic
Keyword(s):  
Recombinant Proteins ◽  
Fusion Proteins ◽  
Kanamycin Resistance ◽  
Expression Vectors ◽  
E Coli ◽  
C Terminus ◽  
N Terminus ◽  
Different Levels ◽  
Streptomyces Tenebrarius

The KgmB methylase from Streptomyces tenebrarius was expressed and purified using the QIAexpress System. Two expression vectors were made: pQEK-N, which places a (His)6 tag at the N-terminus, and pQEK-C, which places a (His)6 tag at the C-terminus of the recombinant KgmB protein. Kanamycin resistance of the E. coli cells containing either the pQEK-N or the pQEK-C recombinant plasmids confirmed the functionality of both KgmB-His fusion proteins in vivo. Interestingly, different levels of expression were observed between these two recombinant proteins. Namely, KgmB methylase with the (His)6 tag at the N-terminus showed a higher level of expression. Purification of the (His)6-tagged proteins using Ni-NTA affinity chromatography was performed under native conditions and the KgmB methylase with (His)6 tag at the N-terminus was purified to homogeneity >95 %. The recombinant KgmB protein was detected on a Western blot using anti-Sgm antibodies.

scholarly journals The multifunctionality of expression systems in Bacillus subtilis: Emerging devices for the production of recombinant proteins

2021 ◽  
pp. 153537022110301
Author(s):  
Caio Coutinho de Souza ◽  
Jander Matos Guimarães ◽  
Soraya dos Santos Pereira ◽  
Luis André Morais Mariúba
Keyword(s):  
Bacillus Subtilis ◽  
Recombinant Proteins ◽  
Large Scale ◽  
Recombinant Expression ◽  
Academic Research ◽  
Future Research ◽  
Expression Systems ◽  
Bioactive Substances ◽  
Exogenous Dna ◽  
Chemical Inducers

Bacillus subtilis is a successful host for producing recombinant proteins. Its GRAS (generally recognized as safe) status and its remarkable innate ability to absorb and incorporate exogenous DNA into its genome make this organism an ideal platform for the heterologous expression of bioactive substances. The factors that corroborate its value can be attributed to the scientific knowledge obtained from decades of study regarding its biology that has fostered the development of several genetic engineering strategies, such as the use of different plasmids, engineering of constitutive or double promoters, chemical inducers, systems of self-inducing expression with or without a secretion system that uses a signal peptide, and so on. Tools that enrich the technological arsenal of this expression platform improve the efficiency and reduce the costs of production of proteins of biotechnological importance. Therefore, this review aims to highlight the major advances involving recombinant expression systems developed in B. subtilis, thus sustaining the generation of knowledge and its application in future research. It was verified that this bacterium is a model in constant demand and studies of the expression of recombinant proteins on a large scale are increasing in number. As such, it represents a powerful bacterial host for academic research and industrial purposes.

scholarly journals A human expression system based on HEK293 for the stable production of recombinant erythropoietin

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Christine Lin Chin ◽  
Justin Bryan Goh ◽  
Harini Srinivasan ◽  
Kaiwen Ivy Liu ◽  
Ali Gowher ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
High Titer ◽  
Expression System ◽  
Mass Spectrometry Analysis ◽  
Hek293 Cells ◽  
Expression Vectors ◽  
Methionine Sulfoximine ◽  
Mammalian Host ◽  
Recombinant Erythropoietin ◽  
Cellular Expression

AbstractMammalian host cell lines are the preferred expression systems for the manufacture of complex therapeutics and recombinant proteins. However, the most utilized mammalian host systems, namely Chinese hamster ovary (CHO), Sp2/0 and NS0 mouse myeloma cells, can produce glycoproteins with non-human glycans that may potentially illicit immunogenic responses. Hence, we developed a fully human expression system based on HEK293 cells for the stable and high titer production of recombinant proteins by first knocking out GLUL (encoding glutamine synthetase) using CRISPR-Cas9 system. Expression vectors using human GLUL as selection marker were then generated, with recombinant human erythropoietin (EPO) as our model protein. Selection was performed using methionine sulfoximine (MSX) to select for high EPO expression cells. EPO production of up to 92700 U/mL of EPO as analyzed by ELISA or 696 mg/L by densitometry was demonstrated in a 2 L stirred-tank fed batch bioreactor. Mass spectrometry analysis revealed that N-glycosylation of the produced EPO was similar to endogenous human proteins and non-human glycan epitopes were not detected. Collectively, our results highlight the use of a human cellular expression system for the high titer and xenogeneic-free production of EPO and possibly other complex recombinant proteins.

Abstract B129: Novel miniprep system for rapid purification of recombinant proteins and antibodies on high capacity membranes

2015 ◽  
Author(s):  
Sayantan Mitra ◽  
Mike Vierra ◽  
Boris Levitan ◽  
Gia Jokadze ◽  
Andrew Farmer
Keyword(s):  
Recombinant Proteins ◽  
High Capacity ◽  
Rapid Purification
Sign in / Sign up

Export Citation Format

Share Document