Method for in vitro protein synthesis

Details Method for in vitro protein synthesis Method for in vitro protein synthesis

1999-01-13

1999-12-07

Lankford, Jr., Leon B.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Enzymatic production of a protein or polypeptide

435 691, 435 701, C12N 500

Patent

active

059981636

ABSTRACT:
A method for performing coupled in vitro transcription and translation reactions is disclosed. In the transcription reaction, the quantity of mRNA produced is limited to a level of less than about 2.5 micrograms in a 10 microliter volume prior to the translation elements being added. Limiting the level of mRNA produced prevents saturation of the translational processes and thus aids in the efficiency and fidelity of the translation process.

REFERENCES:
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patent: 5895753 (1999-04-01), Mierendorf et al.
Coen, et al., "Maize chloroplast DNA fragment encoding the large subunit of ribulosebisphosphate carboxylase," Proc. Natl. Acad. Sci. USA, 74:5487-5491 (1977).
Lewis, et al., "The Origin and Destiny of Adenovirus Proteins," Cold Spring Harbor Symposium on Quantitative Biology, vol. XXXIX p. 581-590 ( ).
Perara & Lingappa, "A Former Amino Terminal Signal Sequence Engineered to an Internal Lcoation Directs Translocation of Both Flanking Protein Domains," The Journal of Cell Biology, 101:2292-2301 (1985).
Stueber, et al., A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences,; IL Press Limited 3134-3148 (1984).
Boehinger Mannheim, "Translation Kit, Reticulocyte, Type II," Cat No. 1103 032 (1989).

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