Cloning and expression of the luxY gene from Vibrio fischeri strain Y-1 in Escherichia coli and complete amino acid sequence of the yellow fluorescent protein

Biochemistry ◽  
1990 ◽  
Vol 29 (23) ◽  
pp. 5509-5515 ◽  
Author(s):  
Thomas O. Baldwin ◽  
Mary L. Treat ◽  
S. Colette Daubner
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Fluorescent Protein ◽  
Vibrio Fischeri ◽  
Yellow Fluorescent Protein ◽  
Cloning And Expression ◽  
Complete Amino Acid Sequence

scholarly journals The cloning and expression of the aroL gene from Escherichia coli K12. Purification and complete amino acid sequence of shikimate kinase II, the aroL-gene product

1986 ◽  
Vol 237 (2) ◽  
pp. 427-437 ◽  
Author(s):  
G Millar ◽  
A Lewendon ◽  
M G Hunter ◽  
J R Coggins
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Transcriptional Start Site ◽  
Cloning And Expression ◽  
Potential Operator ◽  
Gene Encoding ◽  
Shikimate Kinase ◽  
Complete Amino Acid Sequence ◽  
Escherichia Coli K12

The aroL gene encoding the enzyme shikimate kinase II was cloned from Escherichia coli K12. Construction of over-expressing strains permitted for the first time the purification to homogeneity of a monofunctional shikimate kinase. The complete amino acid sequence of shikimate kinase II was determined by a combined nucleotide and direct amino acid sequencing strategy. E. coli shikimate kinase II is a monomeric enzyme containing 173 amino acid residues with a calculated Mr 18,937. The amino acid sequence contains a region homologous with other kinases and ATP-requiring enzymes. Evidence is presented suggesting that the transcriptional start site of the aroL gene is located within a potential operator site.

Determination of the Complete Amino Acid Sequence of Recombinant Human γ-Interferon Produced in Escherichia coli

1986 ◽  
Vol 6 (4) ◽  
pp. 331-336 ◽  
Author(s):  
SHOJIRO YAMAZAKI ◽  
TSUNEO SHIMAZU ◽  
SHIGENOBU KIMURA ◽  
HIROHIKO SHIMIZU
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Complete Amino Acid Sequence ◽  
Γ Interferon

scholarly journals Phenotypic, Biochemical, and Genetic Analysis of KPC-41, a KPC-3 Variant Conferring Resistance to Ceftazidime-Avibactam and Exhibiting Reduced Carbapenemase Activity

2019 ◽  
Vol 63 (12) ◽  
Author(s):  
Linda Mueller ◽  
Amandine Masseron ◽  
Guy Prod’Hom ◽  
Tatiana Galperine ◽  
Gilbert Greub ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Amino Acid ◽  
Klebsiella Pneumoniae ◽  
Genetic Analysis ◽  
Amino Acid Sequence ◽  
Cloning And Expression ◽  
Content Type ◽  
Amino Acid Insertion

ABSTRACT A novel KPC variant, KPC-41, was identified in a Klebsiella pneumoniae clinical isolate from Switzerland. This β-lactamase possessed a 3-amino-acid insertion (Pro-Asn-Lys) located between amino acids 269 and 270 compared to the KPC-3 amino acid sequence. Cloning and expression of the blaKPC-41 gene in Escherichia coli, followed by determination of MIC values and kinetic parameters, showed that KPC-41, compared to those of KPC-3, has an increased affinity to ceftazidime and a decreased sensitivity to avibactam, leading to resistance to ceftazidime-avibactam once produced in K. pneumoniae. Furthermore, KPC-41 exhibited a drastic decrease of its carbapenemase activity. This report highlights that a diversity of KPC variants conferring resistance to ceftazidime-avibactam already circulate in Europe.

scholarly journals Cloning and Expression of Islandisin, a New Thermostable Subtilisin from Fervidobacterium islandicum, in Escherichia coli

2005 ◽  
Vol 71 (7) ◽  
pp. 3951-3958 ◽  
Author(s):  
Carolin Gödde ◽  
Kerstin Sahm ◽  
Stan J. J. Brouns ◽  
Leon D. Kluskens ◽  
John van der Oost ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Serine Protease ◽  
Signal Sequence ◽  
Thermophilic Bacterium ◽  
Cloning And Expression ◽  
Inverse Pcr ◽  
Heat Denaturation ◽  
Catalytic Residues

ABSTRACT A gene encoding a subtilisin-like protease, designated islandisin, from the extremely thermophilic bacterium Fervidobacterium islandicum (DSMZ 5733) was cloned and actively expressed in Escherichia coli. The gene was identified by PCR using degenerated primers based on conserved regions around two of the three catalytic residues (Asp, His, and Ser) of subtilisin-like serine protease-encoding genes. Using inverse PCR regions flanking the catalytic residues, the gene could be cloned. Sequencing revealed an open reading frame of 2,106 bp. The deduced amino acid sequence indicated that the enzyme is synthesized as a proenzyme with a putative signal sequence of 33 amino acids (aa) in length. The mature protein contains the three catalytic residues (Asp177, His215, and Ser391) and has a length of 668 aa. Amino acid sequence comparison and phylogenetic analysis indicated that this enzyme could be classified as a subtilisin-like serine protease in the subgroup of thermitase. The whole gene was amplified by PCR, ligated into pET-15b, and successfully expressed in E. coli BL21(DE3)pLysS. The recombinant islandisin was purified by heat denaturation, followed by hydroxyapatite chromatography. The enzyme is active at a broad range of temperatures (60 to 80°C) and pHs (pH 6 to 8.5) and shows optimal proteolytic activity at 80°C and pH 8.0. Islandisin is resistant to a number of detergents and solvents and shows high thermostability over a long period of time (up to 32 h) at 80°C with a half-life of 4 h at 90°C and 1.5 h at 100°C.

scholarly journals The Complete Amino-Acid Sequence of Elongation Factor Tu from Escherichia coli

1980 ◽  
Vol 108 (2) ◽  
pp. 507-526 ◽  
Author(s):  
Michael D. JONES ◽  
Karen M. NIELSEN ◽  
Brian F. C. CLARK ◽  
Torben E. PETERSEN ◽  
Staffan MAGNUSSON ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Elongation Factor ◽  
Elongation Factor Tu ◽  
Complete Amino Acid Sequence
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