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1、合成生物學(xué)(Synthetic biology)(概念、原理、應(yīng)用),馬飛,人工染色體(技術(shù)),BAC(細(xì)菌人工染色體):Bacteria…以細(xì)菌作為對象,將DNA片段與質(zhì)粒重組后轉(zhuǎn)入細(xì)菌中繁殖YAC(酵母人工染色體):Yeast…以酵母作為對象PAC(噬菌體人工染色體):Phagemid…以噬菌體作為對象TAC(可轉(zhuǎn)化的細(xì)菌人工染色體) MAC(哺乳類人工染色體)…,合成生物學(xué)應(yīng)運(yùn)而生…,Synthetic Biol

2、ogy,What is Synthetic Biology?,Taking an engineering approach to design and applying it to Biology使用工程策略設(shè)計(jì)并應(yīng)用于生物學(xué),What is Synthetic Biology?1. Biology2. Chemistry3. Engineering4. Re-Writing,BiologistsChemistsE

3、ngineers“Re-Writers”,“The code is 3.6 billion years old. It’s time for a re-write.” -Tom Knight,Biology“Test models by building them”,合成生物學(xué),指人們將“基因”連接成網(wǎng)絡(luò),讓細(xì)胞來完成設(shè)計(jì)人員設(shè)想的各種任務(wù)。例如把網(wǎng)絡(luò)同簡單的細(xì)胞相結(jié)合,可提高生物傳感性,幫助檢查人員確定地雷或生物武器的

4、位置。再如向網(wǎng)絡(luò)加入人體細(xì)胞,可以制成用于器官移植的完整器官。,人工合成脊髓灰白質(zhì)炎病毒cDNA,美國紐約大學(xué)Wimmer 實(shí)驗(yàn)室于2002年報(bào)道了化學(xué)合成 脊髓灰白質(zhì)炎病毒cDNA,并用RNA聚合酶將它轉(zhuǎn) 成有感染活力的病毒RNA。開辟了利用已知基因組序列,不需要天然模板,從化合物單體合成感染性病毒的先河。,,Wimmer從裝配平均長度為69 bp的寡核苷酸入手,結(jié)合了化學(xué)合成與無細(xì)胞體系的從頭合成,用了3 年時間完成了這個劃時代

5、的工作。,Venter 實(shí)驗(yàn)室發(fā)展了合成基因組,Φ X-174 噬菌體基因是單鏈環(huán)狀 DNA,是歷史上第一個被純化的DNA 分子,也是第一個被測序的DNA分子。Φ X- 174 噬菌體對動植物無害,是合適的合成研究對象。美國Venter 實(shí)驗(yàn)室發(fā)展了合成基因組的工作, 該實(shí)驗(yàn)室只用兩周就合成了Φ X-174 噬菌體基因 (5,386bp) 。Venter實(shí)驗(yàn)室的技術(shù)改進(jìn)主要有:(1)用凝膠來提純寡核苷酸以減少污染;(2) 嚴(yán)

6、格控制退火連接溫度來防止與不正確的序列發(fā)生連 接;(3)采用聚合酶循環(huán)裝置來裝配連結(jié)產(chǎn)物。,合成生物學(xué)國際會議,2004 年6 月在美國麻省理工學(xué)院舉行了第一屆 合成生物學(xué)國際會議。會上除討論了科學(xué)與技術(shù)問 題外,還討論了合成生物學(xué)當(dāng)前與將來的生物學(xué)風(fēng)險,有關(guān)倫理學(xué)問題,以及知識產(chǎn)權(quán)問題。隨著這個領(lǐng)域的發(fā)展,對于合成生物學(xué)的安全性的考慮愈來愈多?,F(xiàn)在不僅通過合成生成病毒,而且已經(jīng)可以合成細(xì)菌。,合成生物學(xué)開辟了設(shè)計(jì)生命的前景,一

7、方面有可能合成模仿生命物質(zhì)特點(diǎn)的人工化學(xué)系統(tǒng);另一方面也可能重新設(shè)計(jì)微生物如Keasling 實(shí)驗(yàn)室向大腸桿菌中導(dǎo)入青蒿與酵母的基因,使大腸桿菌能在調(diào)節(jié)下合成青蒿素,從而顯示了有效而價廉的治療瘧疾的前景合成生物學(xué)今后將能生成自然界不存在的新的微生物。,應(yīng)用示例,Schultz 實(shí)驗(yàn)室研究向大腸桿菌蛋白質(zhì)生物合成裝置中添入新組份,使之能通過基因生成非天然的氨基酸,結(jié)果取得了成功。但是要在真核細(xì)胞做到這一點(diǎn)還有難度。2003年,Sch

8、ultz 實(shí)驗(yàn)室報(bào)道了一種向酵母加 入非天然氨基酸密碼子的方法,成功地向蛋白質(zhì)中導(dǎo)入了5 種氨基酸。目前,能摻入到蛋白質(zhì)的非天然氨基酸已有80多種。今后將可以直接向蛋白質(zhì)導(dǎo)入順磁標(biāo)記、金屬結(jié)合、光敏異構(gòu)化等的氨基酸,促進(jìn)蛋白質(zhì)結(jié)構(gòu)與功能的研究。,應(yīng)用示例,Brenner 提出向細(xì)胞DNA中摻入天然不存在的堿基來發(fā)展人工遺傳系統(tǒng), 支持人工生命形式。合成生物學(xué)也將對生命起源,其他生命形式的研究作出貢獻(xiàn)。,控制生命,目前,研究人

9、員正在試圖控制細(xì)胞的行為,研制不同的基因線路———即特別設(shè)計(jì)的、相互影響的基因。波士頓大學(xué)生物醫(yī)學(xué)工程師科林斯已研制出一種“套環(huán)開關(guān)”,所選擇的細(xì)胞功能可隨意開關(guān)。加州大學(xué)生物學(xué)和物理學(xué)教授埃羅維茨等人研究出另外一種線路:當(dāng)某種特殊蛋白質(zhì)含量發(fā)生變化時,細(xì)胞能在發(fā)光狀態(tài)和非發(fā)光狀態(tài)之間轉(zhuǎn)換,起到有機(jī)振蕩器的作用,打開了利用生物分子進(jìn)行計(jì)算的大門。,,維斯和加州理工學(xué)院化學(xué)工程師阿諾爾一起,采用“定向進(jìn)化”的方法,精細(xì)調(diào)整研制線路,

10、將基因網(wǎng)絡(luò)插入細(xì)胞內(nèi),有選擇性地促進(jìn)細(xì)胞生長。,發(fā)展方向,維斯目前正在研究另外一群稱為“規(guī)則系統(tǒng)”的基因,他希望細(xì)菌能估計(jì)刺激物的距離,并根據(jù)距離的改變做出反應(yīng)。該項(xiàng)研究可用來探測地雷位置(TNT:生物傳感器)。,,維斯另一項(xiàng)大膽的計(jì)劃是為成年干細(xì)胞編程促進(jìn)某些干細(xì)胞分裂成骨細(xì)胞、肌肉細(xì)胞或軟骨細(xì)胞等,讓細(xì)胞去修補(bǔ)受損的心臟或生產(chǎn)出合成膝關(guān)節(jié)。盡管該工作尚處初級階段,但卻是生物學(xué)調(diào)控領(lǐng)域中重要的進(jìn)展。,J. Craig Vent

11、er:基因組替換,成功利用基因組取代技術(shù),將一種細(xì)菌改變?yōu)榱硪环N與之親緣關(guān)系較為緊密的另一細(xì)菌。這種由J. Craig Venter 進(jìn)行的 “移植(transplantation)”技術(shù),有望將合成基因組插入細(xì)胞,用于生產(chǎn)合成生命。用Mycoplasma mycoides的基因組取代與之關(guān)系密切的 Mycoplasma capricolum的基因組C. Lartigue et al. "Genome transplant

12、ation in bacteria: Changing one species to another" Science, June 28, 2007.,人類歷史上第一個人造染色體合成成功,美科學(xué)家稱“人造生命”技術(shù)已被掌握最具爭議的美國著名科學(xué)家克雷格·文特爾宣布,他的研究小組已經(jīng)合成出人類歷史上首個人造染色體,并有可能創(chuàng)造出首個永久性生命形式,以此作為應(yīng)對疾病和全球變暖的潛在手段。該研究部分由美國能源部出資,希

13、望藉此研制出新型環(huán)保燃料。由文特爾召集,諾貝爾醫(yī)學(xué)獎獲得者漢密爾頓·史密斯領(lǐng)導(dǎo)的研究小組在這方面已經(jīng)進(jìn)行了5年研究。文特爾已用化學(xué)藥品在實(shí)驗(yàn)室中研制出一種合成染色體。,文特爾研究小組研制出的這種新型染色體即實(shí)驗(yàn)室合成支原體(Mycoplasma laboratorium),是一種經(jīng)過簡化拼接的生殖支原體(Mycoplasma genitalium)DNA序列,他們將這種合成支原體移植到活細(xì)胞中,使之在細(xì)胞中起主控作用,變換成

14、一種新的染色體。按照實(shí)驗(yàn)計(jì)劃,最終這個染色體將控制這個細(xì)胞并變成一個新的生命形式。這種新單細(xì)胞生物體被命名為“合成器”,受381個基因控制,包含56萬個堿基對。這些基因是維持細(xì)菌生命所必備的,使它能夠攝食和繁殖。由于新的生物體是在現(xiàn)存生物體上搭建,其繁殖和新陳代謝仍然依賴原來生物體的胞內(nèi)機(jī)制。從這一角度看,它并非完全意義上的新型生命形式。但這種給特定基因賦予特定任務(wù)的觀點(diǎn)已被眾多生物學(xué)家廣泛接受。,“這是人類自然科學(xué)史上一次重

15、大進(jìn)步,顯示人類正在從閱讀基因密碼走向有能力重新編寫密碼,這將賦予科學(xué)家新的能力,從事以前從未做過的研究?!彼M@項(xiàng)突破有助于發(fā)展新能源,應(yīng)對氣候變化造成的負(fù)面影響。如創(chuàng)造出具有特殊功能的新微生物,可被用作替代石油和煤炭的綠色燃料,或用來幫助清除危險化學(xué)物質(zhì)或輻射等;還可用來合成能吸收過多二氧化碳的細(xì)菌,為解決氣候變暖貢獻(xiàn)力量。,然而制造永久生命形式的前景極具爭議性,有可能激起道德、倫理等方面的激烈辯論。加拿大生物倫理學(xué)組織ET

16、C團(tuán)體主任帕特·穆尼說,文特爾制造出了“一個基架,在此基架上人們幾乎可以制造出任何東西”,“它可以用于研究新型藥物,也可以用于對人類產(chǎn)生巨大威脅的生物武器”。,2009:Venter:Science,把蕈狀支原體的基因組加以改造,使它能夠終移植到山羊支原體內(nèi),形成了一個新的蕈狀支原體細(xì)胞。這也是今年這篇科研論文的雛形,在國外的科學(xué)媒體上曾經(jīng)引發(fā)熱烈的討論。,2010年的重要大事:“人造生命”誕生,John Craig Ven

17、ter攪亂了(生命)科學(xué)界,《用化學(xué)合成的基因組構(gòu)建一個細(xì)菌細(xì)胞》,Venter的實(shí)驗(yàn)http://www.science-weekly.cn//skhtmlnews/2010/6/1090.html,實(shí)驗(yàn)對象:蕈狀支原體。支原體是已知的可以自由生活的最小生物,也是最小的原核細(xì)胞。是一種原核微生物, 內(nèi)部結(jié)構(gòu)很簡單,基因組僅有一百多萬堿基對,遠(yuǎn)小于真核生物基因組十億級的堿基數(shù)量,這也是Venter選擇操作它的原因。Venter早

18、在1995年就對生殖支原體測序,并致力于研究維持自由生命的最小基因組。在2008年,Venter的團(tuán)隊(duì)合成了長達(dá)59萬堿基對的生殖支原體基因組。此后,他們選擇生長速度更快的蕈狀支原體來做實(shí)驗(yàn)。如果僅僅從技術(shù)上來說,Venter做了一個無懈可擊的實(shí)驗(yàn),“人造生命”思路和流程都做得無懈可擊。,三個步驟:合成、組裝和移植,合成 :蕈狀支原體的基因組是一條大片段的DNA分子,序列是A、T、G、C四種脫氧核糖核苷酸的排列組合。通過實(shí)驗(yàn)

19、確定維持其生命周期的最小基因組,并加上4個“水印基因”作為標(biāo)記。用計(jì)算機(jī)精確計(jì)算需要合成DNA分子序列,并用化學(xué)方法合成A、T、G、C堿基,并使其按所要求序列延伸。這是它被稱為“人造生命”或者“化學(xué)合成”的關(guān)鍵。Venter用化學(xué)方法合成了一千多個約1kb的DNA片段,作為這次組裝的基本材料。,組裝:因?yàn)楹铣缮飳W(xué)技術(shù)上的局限,不能直接合成上萬堿基對的DNA大分子,所以Venter等人巧妙地借助啤酒酵母和大腸桿菌的幫助,把1Kb

20、的DNA分子有序準(zhǔn)確的連成超過1000kb的片段。移植: Venter等把這個合成基因組移植到不含限制性酶切系統(tǒng)的山羊支原體中,基因組能使用后者的酶系統(tǒng)進(jìn)行自我復(fù)制,經(jīng)過多代繁殖后,長成的菌落已經(jīng)純粹由蕈狀支原體組成。,Venter:“創(chuàng)造了一個計(jì)算機(jī)為父母的生命”,,JCVI:將8個由60個核苷酸組成的DNA片段,首次人工合成實(shí)驗(yàn)老鼠的線粒體基因組,,使用8個只含有60個核苷酸的DNA片段,讓它們同酶和化學(xué)試劑的混合物相結(jié)合,

21、在50℃下孵化1小時,5天內(nèi)合成出了實(shí)驗(yàn)鼠的線粒體基因組,得到的基因組能夠糾正具有線粒體缺陷的細(xì)胞內(nèi)的異常。,用途:生物能源、生物除污…,Venter下一步的計(jì)劃就是合成某種海藻基因組,這種新型海藻可以通過光合作用把空氣中的二氧化碳轉(zhuǎn)化成汽油或者柴油等清潔能源,從而有效解決目前的氣候變化和能源危機(jī)。疫苗、藥物、生物能源、生物除污等,What is Synthetic Biology?,——從原理角度來看,Synthetic Biol

22、ogy,Undergraduates in Synthetic Bio.,international Genetically Engineered Machines,http://parts.mit.edu/registry/index.php/Main_Page,Lego Assembly for DNA Parts,http://parts.mit.edu/registry/index.php/Assembly:Standard_

23、assembly,Self-organized Pattern Formation,What can you make in SB?,Arsenic Detector,膿毒癥,砷,Modifying life,Biotechnology – Techniques that use living organisms or parts of organisms to produce a variety of products (from m

24、edicines to industrial enzymes) Genetic Engineering – Introduction of genetic changes (add, modify, delete) into an organism to achieve some goalSynthetic Biology – Create novel biological functions and tools by modify

25、ing or integrating well-characterized biological components (i.e. genes, promoters) into higher order genetic networks,Synthetic Biology History,1970 – First gene synthesized from scratch (alanine tRNA)1978 – Nobel priz

26、e awarded to Werner Arber, Daniel Nathans and Hamilton Smith for the discovery of restriction enzymes 1978 (Boyer at UCSF) – A synthetic version of the human insulin gene was constructed and inserted into the bacterium

27、E. coli.1980 – Kary Mullis invents PCR1991 – Affymetrix chip-based oligonucleotide synthesis2003 – First iGEM competition, creation of standardized parts libraries at MIT,Biotechnology 1.0 Research Workflow,1. Concept

28、,2. Collect DNA fragments (PCR, isolation, vendors, etc),,,6. Transform,,7. Test,3. Bench work,5. Verify DNA,4. Sequence,,,DNA synthesis costs are dropping,For example the bacteria Mycoplasma genitalium has the smallest

29、genome out of all living cells: 517 genes over 580 kb. Minimal costs of oligo creation (not including error-checking):Mid 1990s: $1/bp = $580,000Circa 2000: $0.35/bp = $203,0002006: $0.11/bp = $63,800Ambitious predi

30、ction of not-too-distant future (Church et al, 2004): $0.00005/bp = $29,Synthesis lengths are increasing,,Commercial DNA Synthesis Companies,,Data Source: Rob Carlson, U of W, Seattle,,BioneerSouth Korea,,CinnagenTehr

31、an, Iran,,Takara BiosciencesDalian, China,,Inqaba BiotecPretoria, South Africa,,FermentasVilnius, Lithuania,,Bio S&T, Alpha DNA,BiocorpMontreal, Canada,,GENEARTRegensberg, Germany,,MWGBangalore, India,,Zelinsk

32、y InstituteMoscow, Russia,,ScinoPharmShan-hua, Taiwan,,GenosphereParis, France,,BiolegioMalden, Netherlands,,AmbionAustin, Texas,,BiosearchNovato, California,,Bio-SynthesisLewisville, Texas,,ChemgenesWilmington,

33、Mass.,,BioSpringFrankfurt am Main, Germany,,BiosourceCamarillo, CA,,DharmaconLafaette, Co.,,CyberGene ABNovum, Sweden,,Cortec DNAKingston, Ontario, CA,,EurogentecBelgium, U.K.,,DNA TechnologyAarhus, Denmark,,Genem

34、ed SynthesisS. San Francisco, CA,,DNA 2.0Menlo Park, CA,,MetabionMunich, Germany,,MicrosynthBalgach, Switzerland,,Japan Bio ServicesJapan,,Blue Heron BiotechnologyBothell, WA,,,GeneworksAdelaide, Australia,,Imperi

35、al Bio-MedicChandigarh, India,,Bioserve BiotechnologiesHyderabad, India,,GenelinkHawthorne, NY.,DNA Synthesis (Caruthers method),Error Rate: 1%0.9950 = 0.60300 seconds per step,Microarray oligonucleotide synthesis,T

36、he power of parallelism,Chip-based versus linear synthesis,Oligonucleotides synthesized,Single-stranded fragments of 50-90nucleotides 3’-overlapping next fragment by 17 nucleotides (Tm calculated 52-56°),Steps 1 t

37、o 5 involve multiple rounds of PCR (heating to 95°, cooling to 56°, and PCR at 72°).Number of rounds depends on number of fragments. Carried out by PCR machine.,Final step of amplification of complete g

38、enedriven by use of excess of terminal single-stranded fragments,PCR-based oligo ligation,In theory, the scale of synthesis is unlimited,Biotechnology 2.0 Research Workflow,,,,,1. Concept,2. Design / debug/ test,4. Des

39、ign oligos,6. Transform,,,7. Test,5. Synthesize DNA,3. Run code,,,What are the implications of DNA synthesis capacity + freedom of information?,The problem: “Dual Use” Research,Dual use research includes life sciences re

40、search:With legitimate scientific purpose That may be misused to pose a biologic threat to public health and/or national security.,How easy is it to get this technology?,What can we do?,Number of Individuals,,Individ

41、ual’s Intent,,,honorable,dishonorable,,,,,Bin Laden Genetics, Inc.,DisgruntledResearcher,Garage Bio-Hacker,BasicResearcher,,Risk spectrum,Basic logic circuits,Borrowing from electrical engineering,Protein Expression B

42、asics,RNA polymerase binds to promoterRNAP transcribes gene into messenger RNARibosome translates messenger RNA into protein,,,Z,,,Z Promoter,Z Gene,,Protein,Transcription,RNA Polymerase,DNA,,Translation,Messenger RNA

43、,Regulation Through Repression and Induction,Repressor proteins can bind to the promoter and block the RNA polymerase from performing transcriptionThe DNA site near the promoter recognized by the repressor is called an

44、operatorThe target gene can code for another repression protein enabling regulatory cascades,,,,,Z Promoter& Operator,Z Gene,R Gene,,R,R,,,,R Promoter,TranscriptionTranslation,DNA Binding,RNA Polymerase,,,Logic Ci

45、rcuits,Proteins are the wires/signalsPromoters + decay implement the gatesAny finite-state digital circuit can be builtFor example, X or Y ? Z,,,,,,,,,X,Y,,,R1,,Z,,R1,R1,,,X,,,,Y,,,,Z,,=,,gene,,gene,,gene,Transcriptio

46、n-Based Inverter,Protein concentrations are analogous to electrical current BUT… proteins do not function in an isolated system and need to be unique,0,,,,1,1,,,,0,,,,,,,,,R,,,R,,Z,,,,Simple Inverter Model,,,,,R,Operato

47、r,Z Gene,Z,,R,,Cooperativity,Cooperative DNA binding is where the binding of one protein increases the likelihood of a second protein bindingCooperativity adds more non-linearity to the systemIncreases switching sensit

48、ivityImproves robustness to noise,,,,,Z Promoter& Operator,Z Gene,R Gene,,R,R,,,,,R Promoter,TranscriptionTranslation,CooperativeDNA Binding,,,RNA Polymerase,R,,Cooperative Inverter Model,,,,,R,R,,Operator,Z Gene

49、,Z,,R,,BioCircuit Computer-Aided Design,SPICE,,BioSPICE,steady state,dynamics,BioSPICE: a prototype biocircuit CAD tool simulates protein and chemical concentrations intracellular circuits, intercellular communication

50、 single cells, small cell aggregates,,Genetic Circuit Elements,,inputmRNA,ribosome,,promoter,,outputmRNA,ribosome,operator,translation,transcription,,RNAp,,RBS,,RBS,,A BioSPICE Inverter Simulation,input,output,,,,repre

51、ssor,,promoter,They work in vivo Flip-flop (Gardner & Collins, 2000)Ring oscillator (Elowitz & Leibler, 2000)However, cells are very complex environmentsCurrent modeling techniques poorly predict behavior,“Pr

52、oof of Concept” Circuits,,time (x100 sec),,,,[A],,,,,[C],[B],,B,_S,_R,,,,,,,,,,,,,,,A,_[R],[B],_[S],[A],time (x100 sec),,time (x100 sec),,,,,RS-Latch (“flip-flop”),Ring oscillator,,Cellular Logic Summary,Current syst

53、ems are limited to less than a dozen gatesThree inverter ring oscillator (Elowitz, 2000)RS latch (Gardner, 2000)Inter-cell communication (Weiss, 2001)A natural repressor-based logic technology presents serious scalab

54、ility issuesScavenging natural repressor proteins is time consumingMatching natural repressor proteins to work together is difficult,Cellular Logic Summary,Sophisticated synthetic biological systems require a scalable

55、cellular logic technology with good cooperativityZinc-finger proteins can be engineered to create many unique proteins relatively easilyZinc-finger proteins can be fused with dimerization domains to increase cooperativ

56、ityA cellular logic technology of only zinc-finger proteins should hopefully be easier to characterize,Single Zinc-Finger Structure,DNA Three BaseRecognition Region,Zinc Atom,,AlphaHelix,,TwoBetaSheets,Poly-Finger Z

57、FPs,A.C. Jamieson, J.C. Miller, and C.O. Pabo. Drug discovery with engineered zinc-finger proteins. Nature Reviews Drug Discovery, May 2003,Complex systems,Q: But if we don’t fully understand all the rules of biology,

58、how can we create anything more than basic systems?A: We can press our limits by modularizing and simplifying as much as possible.,Standardization of ComponentsPredictable performanceOff-the-shelfMechanical Engineer

59、ing (1800s) & the manufacturing revolution (e.g. Henry Ford)AbstractionInsulate relevant characteristics from overwhelming detailSimple components that can be used in combinationFrom Physics to Electrical Engine

60、ering (1900s)Decoupling Design & FabricationRules insulating design process from details of fabricationEnable parts, device, and system designers to work togetherVLSI electronics (1970s),Enabling Synthetic Biolo

61、gy,Characterization,Catalog input-output characteristics of existing and new parts/devices,Standardization,Physical connectionsFunctional connectionsPerformance,SB works via three layers of abstraction,Devices,Parts,Sy

62、stems,,,Abstraction in biology,Devices,Parts,Systems,,,Barriers,- Technological- Legal- Ethical,Synthetic Biology: Intellectual Property,Relationship of synthetic biology to intellectual property law has been largely u

63、nexplored. The relevant research space already contains broad patents on foundational technology. Synthetic biology commons? Tools of open source – property rights coupled with viral licensing,Synthetic Biology: Intell

64、ectual Property,What is patentable and/or copyrightable?Broad biological functions Specific sequences Specific uses Sources of uncertainty in synthetic biology as related to IPR definitionsWhat are effects of altern

65、ate definitions of what is patentable and copyrightable on: Development of field? Efficiency? Justice?,Synthetic Biology: Intellectual Property,Patents on fundamental ideas in synthetic biology Example: A patent on t

66、he idea of a biological part: a piece of DNA with specific function that can be combined with another part in a predefined fashion. Such a patent would be impossible to circumvent. It represents a fundamental concept th

67、at underpins synthetic biology. See Stanford patent on System and method for simulating operation of biochemical systems. United States Patent 5914891,Synthetic Biology: Intellectual Property,Patents on fundamental biol

68、ogical functions Example: A patent on a genetically-encoded inverter Such a patent would be almost impossible to circumvent because it represents a basic biological function that is of use in a range of synthetic biol

69、ogical systems. See US Dept of Health patent on Molecular computing elements, gates and flip-flops. United States Patent 6774222 See Boston University patent on Multi-state genetic oscillator. United States Patent 6737

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