人工合成基因組

1、人工生命artificial life2007年6月 美國科學(xué)家克雷格文特爾和他領(lǐng)導(dǎo)的研究小組宣布，他們首次實(shí)現(xiàn)了完整的基因組在物種間的移植，這一成功為首個(gè)人造生命的降生奏響了序曲。 文特爾說，這次成功讓他向著制造出首個(gè)人造生命又邁進(jìn)一步，他將在幾個(gè)月內(nèi)利用人工合成的基因組展開類似的移植試驗(yàn)，實(shí)現(xiàn)科研史上零的突破。如果試驗(yàn)成功，文特爾就能宣布他造出全球第一個(gè)合成生命形式。2010年5月20日 J. Craig Venter Institute (JCVI), a not-for-profit genomic research organization, published results to

2、day describing the successful construction of the first self-replicating, synthetic bacterial cell. The team synthesized the 1.08 million base pair(1080kb) chromosome of a modified Mycoplasma mycoides genome. The synthetic cell is called Mycoplasma mycoides JCVI-syn1.0 and is the proof of principle

3、that genomes can be designed in the computer, chemically made in the laboratory and transplanted into a recipient cell to produce a new self-replicating cell controlled only by the synthetic genome. This research will be published by Daniel Gibson et al in the May 20th edition of Science Express and

4、 will appear in an upcoming print issue of Science. 1Genome transplantation in bacteria: changing one species to another天然完整基因組種間轉(zhuǎn)移: As a step toward propagation of synthetic genomes, we completely replaced the genome of a bacterial cell with one from another species by transplanting a whole genome

5、as naked DNA. Intact genomic DNA from Mycoplasma mycoides（蕈狀支原體 ） large colony (LC), virtually free of protein, was transplanted into Mycoplasma capricolum（山羊支原體） cells by polyethylene glycol(PEG)-mediated transformation. Cells selected for tetracycline resistance, carried by the M. mycoides LC chro

6、mosome, contain the complete donor genome and are free of detectable recipient genomic sequences. These cells that result from genome transplantation are phenotypically identical to the M. mycoides LC donor strain as judged by several criteria.- Science. 2007 Aug 3; 317:632-82合成基因組-Science論文Science

7、2 July 2010: Vol. 329. no. 5987, pp. 52 - 56 Creation of a Bacterial Cell Controlled by a Chemically Synthesized GenomeDaniel G. Gibson,1 John I. Glass et alWe report the design, synthesis, and assembly of the 1.08megabase pair Mycoplasma mycoides JCVI-syn1.0 genome starting from digitized genome se

8、quence information and its transplantation into a M. capricolum recipient cell to create new M. mycoides cells that are controlled only by the synthetic chromosome. The only DNA in the cells is the designed synthetic DNA sequence, including watermark sequences and other designed gene deletions and p

9、olymorphisms, and mutations acquired during the building process. The new cells have expected phenotypic properties and are capable of continuous self-replication. 3合成生命的基本操作程序4酵母中克隆細(xì)菌全基因組策略Nucleic Acids Research, 2010, Vol. 38, No. 85酵母中克隆支原體全基因組Nucleic Acids Research, 2010, Vol. 38, No. 86合成生命的關(guān)鍵步

10、驟根據(jù)解讀的蕈狀支原體（M. mycoides）基因組DNA順序，文特爾團(tuán)隊(duì)設(shè)計(jì)了1078個(gè)DNA卡盒（cassette）,每個(gè)DNA卡盒長1080bp，其末端有80bp的順序重疊.這些DNA卡盒由JCVI指定的公司lue Heron Biotechnology合成。隨后進(jìn)行三個(gè)操作步驟：第一步，從1078個(gè)DNA卡盒中每次取10個(gè)構(gòu)建了109個(gè)長10kb的DNA卡盒. 第二步，從110個(gè)10kbDNA卡盒中每次取10個(gè)構(gòu)建了11個(gè)長100kb的DNA卡盒.第三步，將11個(gè)長100kb的DNA卡盒在酵母細(xì)胞中彼此連接裝配成完整的人工基因組,全長1080kb,以酵母人工染色體（YAC）的形式擴(kuò)增

11、。在酵母細(xì)胞中人工基因組DNA不發(fā)生甲基化，當(dāng)進(jìn)入受體細(xì)菌后將被受體限制酶降解,因此需要體外甲基化加以保護(hù)。酵母人工染色體（YAC）可以隨同染色體復(fù)制擴(kuò)增。因帶有標(biāo)記基因，可在篩選培養(yǎng)基上穩(wěn)定遺傳。從酵母細(xì)胞中分離擴(kuò)增的人工合成蕈狀支原體基因組DNA，隨后將人工合成基因組導(dǎo)入山羊支原體（Mycoplasma capricolum）受體細(xì)胞，山羊支原體受體細(xì)胞中編碼限制性核酸內(nèi)切酶基因已失活。人工合成的蕈狀支原體基因組DNA在山羊支原體受體細(xì)胞中可以轉(zhuǎn)錄mRNA,并可翻譯成蛋白質(zhì)。轉(zhuǎn)化的山羊支原體受體細(xì)胞基因組DNA或者被蕈狀支原體限制性核酸內(nèi)切酶破壞，或者在細(xì)胞繁殖后丟失。在篩選培養(yǎng)基上，最后

12、只生長人工合成蕈狀支原體基因組的細(xì)胞克隆。最初的實(shí)驗(yàn)中，人工合成的基因組并沒有產(chǎn)上預(yù)想的結(jié)果，沒有細(xì)胞生長。為此文特爾團(tuán)隊(duì)設(shè)計(jì)了一種檢測合成的每個(gè)DNA卡盒中是否存在錯(cuò)誤的方法。他們將天然的11個(gè)100kb的DNA卡盒和人工合成的11個(gè)100 kb的卡盒進(jìn)行搭配重組，用于檢測人工合成的100kb卡盒中是否存在錯(cuò)誤. 凡是檢測到功能有缺陷的人工卡盒，隨即進(jìn)行DNA測序，找出存在的突變并給予矯正, 確保進(jìn)入人工基因組合成的卡盒完全正確。First Self-Replicating Synthetic Bacterial Cell 20-May-2010 7合成生命操作圖解創(chuàng)造這個(gè)可復(fù)制的試驗(yàn)性單細(xì)

13、胞生物花費(fèi)了4,000萬美元 取名 Synthia:人造兒 8合成基因組結(jié)構(gòu)圖9人工合成基因組中的水印 The secret amino acid messages contained in watermarks that were embedded in the worlds first manmade bacterial genome.NCBI checked into the genetic sequence submitted by Venters Institute and found the watermarks hidden in plain sight. The five coded mess