93 research outputs found

    Expression Screening of Fusion Partners from an E. coli Genome for Soluble Expression of Recombinant Proteins in a Cell-Free Protein Synthesis System

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    While access to soluble recombinant proteins is essential for a number of proteome studies, preparation of purified functional proteins is often limited by the protein solubility. In this study, potent solubility-enhancing fusion partners were screened from the repertoire of endogenous E. coli proteins. Based on the presumed correlation between the intracellular abundance and folding efficiency of proteins, PCR-amplified ORFs of a series of highly abundant E. coli proteins were fused with aggregation-prone heterologous proteins and then directly expressed for quantitative estimation of the expression efficiency of soluble translation products. Through two-step screening procedures involving the expression of 552 fusion constructs targeted against a series of cytokine proteins, we were able to discover a number of endogenous E. coli proteins that dramatically enhanced the soluble expression of the target proteins. This strategy of cell-free expression screening can be extended to quantitative, global analysis of genomic resources for various purposes.National Research Foundation of KoreaKorea (South). Ministry of Education, Science and Technology (MEST) (grant 2011K000841)Korea (South). Ministry of Education, Science and Technology (MEST) (grant 2011-0027901

    Fertility Regulation

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    In the past two centuries the proportion of couples using some form of conscious pregnancy-prevention has risen from close to zero to about two-thirds. In European populations this radical change in behaviour occurred largely between 1870 and 1930 without the benefit of highly effective methods. In Asia, Africa and Latin America, the change took place after 1950 since when the global fertility rate has halved from 5.0 births to 2.5 births per woman. In this chapter we describe the controversies surrounding the idea of birth control and the role of early pioneers such as Margaret Sanger; the advances in contraceptive and abortion technologies; the ways in which family planning has been promoted by many governments, particularly in Asia; trends in use of specific methods; the problems of discontinuation of use; and the incidence of unintended pregnancies and abortions

    A Phenomenological Approach to Donnie Darko

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