Premium
Estimation of the Rate of Cloned Gene Integration via Retrotransposition
Author(s) -
Wang Xiaohai,
Da Silva Nancy A.
Publication year - 1997
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp9700061
Subject(s) - retrotransposon , gene , saccharomyces cerevisiae , genetics , biology , population , yeast , heterologous , genome , transposable element , demography , sociology
A theoretical method has been developed to estimate retrotransposition (integration) rates of the Saccharomyces cerevisiae Ty3 and Ty1 retrotransposons bearing heterologous and homologous genes ( neo r , HIS3 ) . The method is based on population growth modeling and Lea and Coulson's maximal likelihood method for mutation rate estimation (Lea and Coulson, 1949) . This method has allowed us to examine directly retrotransposition rates of GAL ‐regulated marked Ty3 and Ty1 elements into the whole yeast genome, not just a particular DNA sequence, for the purpose of cloned gene integration. The integration rates of a Ty3‐ neo r system (ca. 1 × 10 −3 cell −1 generation −1 ) and a Ty1‐ neo r system (ca. (2–3) × 10 −3 cell −1 generation −1 ) were not significantly affected by temperature (18 and 30 °C) . However, the retrotransposition rate of the Ty3‐ neo r ‐ HIS3 system increased from ca. 2 × 10 −5 to 2 × 10 −4 cell −1 generation −1 when the temperature was decreased from 30 to 18 °C. The retrotransposition rate of Ty3‐ neo r was significantly higher than that of Ty3‐ neo r ‐ HIS3 and slightly lower than that of Ty1‐ neo r . This method can be used to estimate integration rates of other Ty3 and Ty1 elements and to evaluate the efficiency of Ty‐mediated cloned gene integration.