Mutagenesis in Vitro

Stability and Purification of the Recombinant Protein. There are no hard and fast rules specifying, eg, whether a recombinant protein is available in a soluble state in the cell. In some cases, the expression system must be engineered by in vitro mutagenesis to optimize overall yield of the protein. 

BioRad product bulletin, Lit-245 0890 90-0910a, Muta-Gene M13 In Vitro Mutagenesis Kit, Cat. No. 170-3580, BioRad Laboratories, Hercules, Ca., 1990. 

Yang, J. and Kramer, J.M. (1994) In vitro mutagenesis of Caenorhabditis elegans cuticle collagens identifies a potential subtilisin-like protease cleavage site and demonstrates that carboxyl domain disulfide bonding is required for normal function but not assembly. Molecular and Cellular Biology 14, 2722-2730. 

Savarese TM, Fraser CM. In vitro mutagenesis and the search for structure-function relationships among G protein-coupled receptors. Biochem J 1992 283 1-19. 

Sullivan, M. L. and Vierstra, R. D. Cloning of a 16-kDa ubiquitin carrier protein from wheat and Arabidopsis thaliana. Identification of functional domains by in vitro mutagenesis. J. Biol. Chem. 1991, 266, 23878-85. 

Zamocky M, Koller F. 1999. Understanding the structure and function of catalases clues from molecular evolution and in vitro mutagenesis. Prog Biophys Mol Biol 72 19-66. 

Darroudi, F. Natarajan, A.T. (1993) Metabolic activation of chemicals to mutagenic carcinogens by human hepatoma microsomal extracts in Chinese hamster ovary cells (in vitro). Mutagenesis, 8, 11-15... 

DNA sequence information is the starting point for other applications, including the expression of a gene product, the search for related sequences in biological samples, in vitro mutagenesis of the sequence, and structure—function studies of gene expression. 

J. Braman, In Vitro Mutagenesis Protocols, second edition, Humana Press, Totowa, NJ... 

The process of selecting an RNA catalyst with a particular function begins with a large pool of random sequences. This pool can be acquired by producing random single-stranded DNA on a nucleic acid synthesizer or by in vitro mutagenesis of an existing nucleic acid library [18,19]. Random sequence pools produced by chemical... 

Dieldrin has been undergoing in vitro mutagenesis/genetic toxicity testing (CH0 cells) to have been completed in 1985. Testing on endrin for mammalian mutagenesis/genetic toxicity (mouse lymphoma cells) was to have started in 1985 (ref. 89, pp. 37 and 49). 

Evaluation of the Active Site Residues in AspAT by in vitro Mutagenesis... 

In vitro mutagenesis is used to evaluate how the individual residues are involved in enzyme catalysis this is difficult to pursue with traditional methods, such as physicochemical or chemical modification. Mutagenesis study has been successfully employed to define the role of active-site residues in AspAT. It was found that some of the active-site residues are involved in coenzyme binding, some are used for substrate recognition, and others are involved directly in catalysis. 

Heffron, F., So, M., and McCarthy, B. J. (1978). In vitro mutagenesis of a circular DNA molecule by using synthetic restriction sites. Proc. Natl. Acad. Sci. USA, 75, 6012-6016. 

Directed evolution consists of cycles of diversification and selection. Because ribosome display takes place entirely in vitro, it can ideally be combined with in vitro methods of generating sequence diversity. Since true evolution requires diversification in each cycle, this facile alternation between in vitro mutagenesis and in vitro selection is one of the attractions of the ribosome display method. 

Messing, J.B., 1981. M13tnp2 and derivatives a molecular cloning system for DNA sequencing, strand specific hybridization and in vitro mutagenesis, in Third Cleveland Symposium on Macromolecular Recombinant DNA, Ed. A. Walton (Elsevier, Amsterdam) 143-153. 

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