The N-Terminal Signal Sequence and the Last 98 Amino Acids Are Not Essential for the Secretion of Bacillus sp. TS-23 α-Amylase in Escherichia coli

2001 ◽  
Vol 43 (3) ◽  
pp. 170-175 ◽  
Author(s):  
Huei-Fen Lo ◽  
Long-Liu Lin ◽  
Chien-Cheng Li ◽  
Wen-Hwei Hsu ◽  
Chen-Tien Chang
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Signal Sequence ◽  
Bacillus Sp

scholarly journals Molecular cloning and expression in Escherichia coli of a Trichoderma viride endo-beta-(1 6)-galactanase gene

2004 ◽  
Vol 377 (3) ◽  
pp. 749-755 ◽  
Author(s):  
Toshihisa KOTAKE ◽  
Satoshi KANEKO ◽  
Aya KUBOMOTO ◽  
Md. Ashraful HAQUE ◽  
Hideyuki KOBAYASHI ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Signal Sequence ◽  
Trichoderma Viride ◽  
Degree Of Polymerization ◽  
Glycoside Hydrolases ◽  
Cloning And Expression ◽  
Arabinogalactan Protein ◽  
Reading Frame ◽  
Cloned Gene

A gene encoding endo-β-(1→6)-galactanase from Trichoderma viride was cloned by reverse transcriptase–PCR and expressed in Escherichia coli. The gene contained an open reading frame consisting of 1437 bp (479 amino acids). The deduced amino acid sequence of the protein showed little similarity with other known glycoside hydrolases. A signal sequence (20 amino acids) was found at the N-terminal region of the protein and the molecular mass of the mature form was calculated to be 50.488 kDa. The gene product expressed in E. coli as a recombinant protein fused with thioredoxin and His6 tags had almost the same substrate specificity and mode of action as native enzyme purified from a commercial cellulase preparation of T. viride, i.e. recombinant enzyme endo-hydrolysed β-(1→6)-galacto-oligomers with a DP (degree of polymerization) higher than 3, and it could also hydrolyse α-l-arabinofuranosidase-treated arabinogalactan protein from radish. It produced β-(1→6)-galacto-oligomers ranging from DP 2 to at least 8 at the initial hydrolysis stage and galactose and β-(1→6)-galactobiose as the major products at the final reaction stage. These results indicate that the cloned gene encodes an endo-β-(1→6)-galactanase. As far as we know, this is the first time an endo-β-(1→6)-galactanase has been cloned.

scholarly journals Characterization and in Vivo Cloning of prlC, a Suppressor of Signal Sequence Mutations in Escherichia coli K12

Genetics ◽  
1987 ◽  
Vol 116 (4) ◽  
pp. 513-521
Author(s):  
Nancy J Trun ◽  
Thomas J Silhavy
Keyword(s):  
Escherichia Coli ◽  
Genetic Mapping ◽  
Signal Sequence ◽  
Illegitimate Recombination ◽  
Mutant Phenotype ◽  
E Coli ◽  
Physical Location ◽  
Sequence Deletion ◽  
New Alleles

ABSTRACT The prlC gene of E. coli was originally identified as an allele, prlC1, which suppresses certain signal sequence mutations in the genes for several exported proteins. We have isolated six new alleles of prlC that also confer this phenotype. These mutations can be placed into three classes based on the degree to which they suppress the lamBsignal sequence deletion, lamBs78. Genetic mapping reveals that the physical location of the mutations in prlC correlates with the strength of the suppression, suggesting that different regions of the gene can be altered to yield a suppressor phenotype. We also describe an in vivo cloning procedure using λplacMu9H. The procedure relies on transposition and illegitimate recombination to generate a specialized transducing phage that carries prlC1. This method should be applicable to any gene for which there is a mutant phenotype.

scholarly journals ANTIPODAL SPECIFICITY IN THE INHIBITION OF GROWTH OF ESCHERICHIA COLI BY AMINO ACIDS

1948 ◽  
Vol 174 (2) ◽  
pp. 391-398
Author(s):  
Yutaka. Kobayashi ◽  
Marguerite. Fling ◽  
Sidney W. Fox
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Inhibition Of Growth

scholarly journals A novel chitosanase from Bacillus sp. Strain K17: Gene cloning and expression in Escherichia coli

2002 ◽  
Vol 2 (1) ◽  
pp. 227-228 ◽  
Author(s):  
R. Yatsunami ◽  
Y. Sakihama ◽  
M. Suzuki ◽  
T. Fukazawa ◽  
S. Shimizu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gene Cloning ◽  
Cloning And Expression ◽  
Gene Cloning And Expression ◽  
Bacillus Sp ◽  
Expression In Escherichia Coli
Sign in / Sign up

Export Citation Format

Share Document