In vivo import of yeast adenylate kinase into mitochondria affected by site-directed mutagenesis

FEBS Letters ◽  
1992 ◽  
Vol 299 (3) ◽  
pp. 267-272 ◽  
Author(s):  
Viktor Magdolen ◽  
Roland Schricker ◽  
Gertrud Strobel ◽  
Herbert Germaier ◽  
Wolfhard Bandlow
Keyword(s):  
Adenylate Kinase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis

scholarly journals In vivo elimination of parental clones in general and site-directed mutagenesis

2015 ◽  
Vol 417 ◽  
pp. 67-75 ◽  
Author(s):  
Erika G. Holland ◽  
Felicity E. Acca ◽  
Kristina M. Belanger ◽  
Mary E. Bylo ◽  
Brian K. Kay ◽  
...  
Keyword(s):  
Site Directed Mutagenesis ◽  
Directed Mutagenesis

scholarly journals The consequences of deglycosylation of recombinant intra-melanosomal domain of human tyrosinase

2017 ◽  
Vol 399 (1) ◽  
pp. 73-77 ◽  
Author(s):  
Monika B. Dolinska ◽  
Yuri V. Sergeev
Keyword(s):  
Oculocutaneous Albinism ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Multiple Site ◽  
Insect Larvae ◽  
Melanin Biosynthesis ◽  
Human Tyrosinase

AbstractTyrosinase, a melanosomal glycoenzyme, catalyzes initial steps of the melanin biosynthesis. While glycosylation was previously studiedin vivo, we present three recombinant mutant variants of human tyrosinase, which were obtained using multiple site-directed mutagenesis, expressed in insect larvae, purified and characterized biochemically. The mutagenesis demonstrated the reduced protein expression and enzymatic activity due to possible loss of protein stability and protein degradation. However, the complete deglycosylation of asparagine residuesin vitro, including the residue in position 371, interrupts tyrosinase function, which is consistent with a melanin loss in oculocutaneous albinism type 1 (OCA1) patients.

An Efficient Site-Directed Mutagenesis Method In Vivo in Escherichia Coli

2012 ◽  
Vol 195-196 ◽  
pp. 407-411
Author(s):  
Mu Qing Qiu
Keyword(s):  
Escherichia Coli ◽  
Homologous Recombination ◽  
Recombinant Plasmid ◽  
Genomic Dna ◽  
Gene Replacement ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis

In order to develop an efficient site-directed mutagenesis method in vivo, the tests were tested by the following methods. The methods that the fragment knockouted ompR gene was constructed through overlapping PCR, digested by Notand Sal, ligated to plasmid pKOV were applied. The recombination plasmid was transformed into Escherichia coli WMC-001 strain, integrated into the genomic DNA through two step homologous recombination. The Escherichia coli WMC-001/ompR-mutant was obtained due to gene replacement. The fragment of the mutant ompR gene was amplified through overlapping PCR, cloned into pKOV vector. The recombinant plasmid was introduced into Escherichia coli WMC-001/ompR-mutant. The Escherichia coli WMC-001/ompR mutant was also obtained due to gene replacement. Results: The site-directed mutagenesis has been successfully constructed in the ompR gene by sequencing. Conclusion: The method is effective for construction of gene site-directed mutagenesis in vivo.

Steady-State Kinetics of Thr35 and Thr39 Mutants in Human Adenylate Kinase by Site-Directed Mutagenesis

1996 ◽  
Vol 49 (5-6) ◽  
pp. 305-312
Author(s):  
Takanori Ayabe ◽  
Hitoshi Takenaka ◽  
Toshio Onitsuka ◽  
Koichiro Shibata ◽  
Osamu Takenaka ◽  
...  
Keyword(s):  
Steady State ◽  
Adenylate Kinase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Steady State Kinetics ◽  
Kinetics Of

scholarly journals Catalytic roles of lysines (K9, K27, K31) in the N-terminal domain in human adenylate kinase by random site-directed mutagenesis

IUBMB Life ◽  
1996 ◽  
Vol 40 (5) ◽  
pp. 897-906
Author(s):  
Takanori Ayabe ◽  
Seung Kyu Park ◽  
Hitoshi Takenaka ◽  
Michihiro Sumida ◽  
Seiichi Uesugi ◽  
...  
Keyword(s):  
Adenylate Kinase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Terminal Domain ◽  
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