Expression of the N2 fixation gene operon of Paenibacillus sp. WLY78 under the control of the T7 promoter in Escherichia coli BL21

2015 ◽  
Vol 37 (10) ◽  
pp. 1999-2004 ◽  
Author(s):  
Lihong Zhang ◽  
Xiaomeng Liu ◽  
Xinxin Li ◽  
Sanfeng Chen
Keyword(s):  
Escherichia Coli ◽  
N2 Fixation ◽  
T7 Promoter ◽  
Escherichia Coli Bl21 ◽  
Paenibacillus Sp

scholarly journals Isolation of a Gene Responsible for the Oxidation oftrans-Anethole topara-Anisaldehyde by Pseudomonas putida JYR-1 and Its Expression in Escherichia coli

2012 ◽  
Vol 78 (15) ◽  
pp. 5238-5246 ◽  
Author(s):  
Dongfei Han ◽  
Ji-Young Ryu ◽  
Robert A. Kanaly ◽  
Hor-Gil Hur
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Putida ◽  
Hypothetical Protein ◽  
Aldehyde Group ◽  
Agrobacterium Vitis ◽  
T7 Promoter ◽  
Content Type ◽  
E Coli ◽  
Cell Extracts ◽  
Isoeugenol Monooxygenase

ABSTRACTA plasmid, pTA163, inEscherichia colicontained an approximately 34-kb gene fragment fromPseudomonas putidaJYR-1 that included the genes responsible for the metabolism oftrans-anethole to protocatechuic acid. Three Tn5-disrupted open reading frame 10 (ORF 10) mutants of plasmid pTA163 lost their abilities to catalyzetrans-anethole. Heterologously expressed ORF 10 (1,047 nucleotides [nt]) under a T7 promoter inE. colicatalyzed oxidative cleavage of a propenyl group oftrans-anethole to an aldehyde group, resulting in the production ofpara-anisaldehyde, and this gene was designatedtao(trans-anetholeoxygenase). The deduced amino acid sequence of TAO had the highest identity (34%) to a hypothetical protein ofAgrobacterium vitisS4 and likely contained a flavin-binding site. Preferred incorporation of an oxygen molecule from water intop-anisaldehyde using18O-labeling experiments indicated stereo preference of TAO for hydrolysis of the epoxide group. Interestingly, unlike the narrow substrate range of isoeugenol monooxygenase fromPseudomonas putidaIE27 andPseudomonas nitroreducensJin1, TAO fromP. putidaJYR-1 catalyzed isoeugenol,O-methyl isoeugenol, and isosafrole, all of which contain the 2-propenyl functional group on the aromatic ring structure. Addition of NAD(P)H to the ultrafiltered cell extracts ofE. coli(pTA163) increased the activity of TAO. Due to the relaxed substrate range of TAO, it may be utilized for the production of various fragrance compounds from plant phenylpropanoids in the future.

Enzyme replacement therapy for Morquio A: an active recombinant N-acetylgalactosamine-6-sulfate sulfatase produced in Escherichia coli BL21

2010 ◽  
Vol 37 (11) ◽  
pp. 1193-1201 ◽  
Author(s):  
Alexander Rodríguez ◽  
Ángela J. Espejo ◽  
Alejandra Hernández ◽  
Olga L. Velásquez ◽  
Lina M. Lizaraso ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Enzyme Replacement Therapy ◽  
Replacement Therapy ◽  
Escherichia Coli Bl21 ◽  
Enzyme Replacement ◽  
Morquio A

Ethanol effects on the overexpression of heterologous catalase in Escherichia coli BL21 (DE3)

2018 ◽  
Vol 103 (3) ◽  
pp. 1441-1453 ◽  
Author(s):  
Hongchen Zheng ◽  
Zhenxiao Yu ◽  
Wenju Shu ◽  
Xiaoping Fu ◽  
Xingya Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Escherichia Coli Bl21

Overexpression of a plant cysteine synthase gene and biosynthesis of a plant specific metabolite, β-(pyrazol-1-yl)-L-alanine, in Escherichia coli

1994 ◽  
Vol 72 (1) ◽  
pp. 188-192 ◽  
Author(s):  
Kazuki Saito ◽  
Reiko Kanda ◽  
Makoto Kurosawa ◽  
Isamu Murakoshi
Keyword(s):  
Escherichia Coli ◽  
Transcriptional Control ◽  
Spinacia Oleracea ◽  
Higher Plants ◽  
Total Soluble Protein ◽  
T7 Promoter ◽  
Cysteine Synthase ◽  
Mechanistic Investigation

Cysteine synthase (EC 4.2.99.8) in higher plants is responsible for biosynthesis of not only cysteine but also some nonprotein amino acids such as β-(pyrazol-1-yl)-L-alanine. The cDNA of a cysteine synthase from spinach (Spinacia oleracea) was inserted into pET8c (=pET3d) under the transcriptional control of strong T7 promoter to yield an overexpression vector pCEK1. The amount of the exogenous cysteine synthase was increased up to 40% of the total soluble protein of Escherichia coli transformed with pCEK1. β-(Pyrazol-1-yl)-L-alanine, a specific metabolite in plants of the Cucurbitaceae, was biosynthesized by overexpressed cysteine synthase from pyrazole in the presence of O-acetyl-L-serine and serine, in vitro and in vivo, respectively. The present study provides the system for mechanistic investigation of biosynthesis of cysteine and biogenetically related β-substituted alanines at molecular genetic level.

Characterization of the T7 promoter system for expressing penicillin acylase in Escherichia coli

2006 ◽  
Vol 72 (3) ◽  
pp. 529-536 ◽  
Author(s):  
Yali Xu ◽  
Stefan Rosenkranz ◽  
Chiao-Ling Weng ◽  
Jeno M. Scharer ◽  
Murray Moo-Young ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Penicillin Acylase ◽  
T7 Promoter ◽  
Promoter System

Analysis of p53 expression in human tumours: an antibody raised against human p53 expressed in Escherichia coli

1992 ◽  
Vol 101 (1) ◽  
pp. 183-189 ◽  
Author(s):  
C.A. Midgley ◽  
C.J. Fisher ◽  
J. Bartek ◽  
B. Vojtesek ◽  
D. Lane ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polyclonal Antibodies ◽  
P53 Protein ◽  
Expression System ◽  
High Titre ◽  
T7 Promoter ◽  
Aberrant Expression ◽  
P53 Expression ◽  
Normal Human ◽  
Very High

A cDNA encoding the complete normal human p53 protein was expressed in Escherichia coli using an expression system based on the bacteriophage T7 promoter. The cDNA was adapted so that the full-length protein was produced without fusion to any other sequence. Large amounts of the protein were isolated and the purified protein used to produce very high titre polyclonal antibodies to p53. These new antibodies permit the sensitive detection of p53 and p53 complexes in ELISA and immunoblotting assays. Most importantly, they also permit the detection of p53 in archival tumour material that has been conventionally fixed in formalin and embedded in paraffin wax. Using this reagent we have found that aberrant expression of p53 is a frequent feature of human breast cancer. We are able to recognise six different classes of p53 expression pattern that may be of help in the subclassification of breast tumours.

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