基因與基因組的精細(xì)合成原理和技術(shù)研究報(bào)告

趙國(guó)屏　王金　周湘元

摘 要：合成生物學(xué)無(wú)論是對(duì)于理解生命的本質(zhì)還是對(duì)發(fā)展改造生命的技術(shù)手段都具有非常重要的作用。在聚合酶拼接法（PCA）技術(shù)發(fā)明之后，隨著大量設(shè)計(jì)引物和DNA測(cè)序的高效率和低成本，特別是大片段DNA的拼接、克隆等多項(xiàng)技術(shù)的建立和發(fā)展，使得從頭合成大片段DNA甚至的微生物基因組成為現(xiàn)實(shí)。然而，作為一個(gè)新興的領(lǐng)域，合成生物學(xué)仍面臨著諸多的問(wèn)題，無(wú)論是基礎(chǔ)理論還是工程技術(shù)方面都還有待進(jìn)一步的發(fā)展和完善。該課題重點(diǎn)發(fā)展“自下而上”的使能技術(shù)，包括DNA高通量、高保真合成和大片段DNA高效拼接。此外，亦進(jìn)行染色體的改造來(lái)為天然產(chǎn)物的高效表達(dá)發(fā)展優(yōu)化的底盤(pán)細(xì)胞。

關(guān)鍵詞：DNA合成 DNA拼接 高通量 高保真 大片段

Abstract：Synthetic biology is very important both for the understanding of the nature of life and for the development of new techniques. After the invention of polymerase chain assembly （PCA）， the much lower cost and higher efficiency in primer design and DNA sequencing and in particular the establishment and development of new technologies for splicing， cloning，etc.， make the de novo synthesis of large DNA fragment or even microbial genome a reality. However， as an emerging field， synthetic biology is still facing a lot of problems， among which both the theory and engineering technology are to be further developed and improved. In this project， “bottom-up” enabling technologies， including those for high-throughput， high-fidelity DNA synthesis and efficient assembly of large DNA fragments. In addition， optimized chassis cells are constructed through directly editing the chromosome， which can be used for efficient expression of natural products.

Key Words：DNA synthesis；DNA assembly；High throughput；High fidelity；Large DNA fragments

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