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Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system.

ABSTRACT: Replication of all living cells relies on the multirounds flow of the central dogma. Especially, expression of DNA replication proteins is a key step to circulate the processes of the central dogma. Here we achieved the entire sequential transcription-translation-replication process by autonomous expression of chromosomal DNA replication machineries from a reconstituted transcription-translation system (PURE system). We found that low temperature is essential to express a complex protein, DNA polymerase III, in a single tube using the PURE system. Addition of the 13 genes, encoding initiator, DNA helicase, helicase loader, RNA primase and DNA polymerase III to the PURE system gave rise to a DNA replication system by a coupling manner. An artificial genetic circuit demonstrated that the DNA produced as a result of the replication is able to provide genetic information for proteins, indicating the in vitro central dogma can sequentially undergo two rounds.

SUBMITTER: Fujiwara K

PROVIDER: S-EPMC3737561 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

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Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system.

Fujiwara Kei K Katayama Tsutomu T Nomura Shin-Ichiro M SM

Nucleic acids research 20130603 14

Replication of all living cells relies on the multirounds flow of the central dogma. Especially, expression of DNA replication proteins is a key step to circulate the processes of the central dogma. Here we achieved the entire sequential transcription-translation-replication process by autonomous expression of chromosomal DNA replication machineries from a reconstituted transcription-translation system (PURE system). We found that low temperature is essential to express a complex protein, DNA po ...[more]

PMID: 23737447

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Project description:BackgroundTo prove the possibility of implementing the New Rural Cooperative Medical System (NRCMS) at different levels with a premium funding according to their economic level in developed and less developed areas in Guangdong province, and study the insurable inpatients in different types of regions, taking into account limitations of indemnities and loss ratios.MethodAll data samples were randomly collected from the NRCMS Department, Guangdong Province. Gross domestic product (GDP) at 10000 Yuan per capita was employed to divide Guangdong into two economic levels: (1) economically developed & (2) less economically developed regions. A descriptive analysis about tendency of raising premium and reimbursement ratios of common fund was performed with independent samples and t-test as well as implementing a model to evaluate the differences in premium contribution differences in co-payments, thresholds, and rebates. Also, a qualitative study measured several economic factors to evaluate farmers' financial and social potency in contributing to the NRCMS.ResultA higher GDP per capita were found within economically developed regions (p < 0.05) than in less developed areas, with higher tendency for funding capacity and average funding capability in villages and towns within economically developed regions (p < 0.05) than in economically less developed. Maximum benefits between two regions in medical insurance coverage showed significant difference (p < 0.05); differences between basic medical insurance coverage between two regions was insignificant (p > 0.05); nevertheless, economically developed regions showed higher threshold and rebates with less co-payments in the economically developed than less developed.ConclusionDespite some loop holes in the NRCMS, the system is workable, but needs more strengthening by encouraging farmers' participation into NRCMS with a necessity to implement a new reimbursement payment system by health care providers. In addition it is proposed that for maximum benefits another premium funding should be secured.

Dataset Information

Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system.

Publications

Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system.

Similar Datasets

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