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  • TS. CHU HONG MU

    C S V PHNG PHP SINH HC PHN T

    NH XUT BN I HC S PHM

  • Li ni u.................................................................................................................................4

    Chng 1: H GENE..............................................................................................................5

    1. KHI NIM H GENE (GENOME) ..................................................................5 2. AXIT NUCLEIC ..................................................................................................9 3. ADN V TI BN ADN..................................................................................11 4. ARN V C CH PHIN M .........................................................................19 THO LUN...........................................................................................................23

    Chng 2: C IM CU TRC GENE CA SINH VT PROKARYOT V EUKARYOT ...........................................................................................................25

    1. C IM CU TRC GENE PROKARYOT ..........................................25 2. CU TRC PHN ON GENE EUKARYOT .........................................25 3. MT S TRNH T ADN................................................................................27 THO LUN...........................................................................................................32

    Chng 3: MI LIN H GIA ADN, ARN, PROTEIN................................................33

    1. THNG TIN DI TRUYN V MT M DI TRUYN .................................33 2. PROTEIN ...........................................................................................................39 3. QU TRNH SINH TNG HP PROTEIN.....................................................42 4. IU HO BIU HIN GENE ........................................................................45

    THO LUN......................................................................................................................49

    Chng 4: ENZYME S DNG TRONG K THUT SINH HC PHN T ............50

    1. ENYME GII HN (RESTRICTION ENZYME HOC ENZYME CT HN CH (RESTRICTION ENDONUCLEASE - RE) .............................................50

    2. CC NHM ENZYME KHC.........................................................................57 3. ENZYME NUCLEASE......................................................................................60

    THO LUN......................................................................................................................61

    Chng 5: XC NH MI LIN QUAN GIA PROTEIN V C TNH SINH VT ...............................................................................................................62

    1. MT S LOI PROTEIN CHC NNG........................................................62 2. QUY TRNH XC NH CH TH PHN T PROTEIN..............................66 3. K THUT PHN TCH PROTEIN .................................................................67 4. K THUT PHN TCH ENZYME..................................................................74 5. NGHIN CU RIPS BNG WESTERN BLOT ..............................................77

    THO LUN......................................................................................................................78

    Chng 6: K THUT SINH HC PHN T TRONG PHN TCH H GENE SINH VT..........................................................................................................................79

    1. PHNG PHP TCH CHIT ADN..............................................................79 2. LAI PHN T ...................................................................................................83 3. XC NH TRNH T NUCLEOTIT CA ADN ..........................................85 4. RFLP TECHNOLOGY (K thut phn tch hin tng a hnh ca di cc

    phn on ADN).................................................................................................87

    2

  • THO LUN......................................................................................................................91

    Chng 7: PHN NG CHUI POLIMERASE (POLIMERASE CHAIN REACTION - PCR)................................................................................................92

    1. PHN NG CHUI POLIMERASE (PCR) ....................................................92 2. PHN TCH TNH A HNH CA ADN C NHN BN NGU

    NHIN (RANDQM AMPLIFIED PQLIMORPHIC DAN - RAPD) ................99 4. PHN TCH TNH A HNH CHIU DI CC PHN ON ADN C

    NHN BN C CHN LC .........................................................................106 THO LUN.........................................................................................................108

    Chng 8: T BO CH V VECTOR ..........................................................................109

    1. T BO CH ..................................................................................................109 2. VECTOR ..........................................................................................................111 3. PHNG PHP CHUYN VECTOR TI T HP VO T BO CH ..122 THO LUN.........................................................................................................125

    Chng 9: PHN LP GENE TCH DNG PHN T V BIU HIN GENE......126

    1. PHN LP GENE ...........................................................................................126 2. TCH DNG GENE .......................................................................................127 3. BIU HIN GENE...........................................................................................139 THO LUN.........................................................................................................151 Ti liu tham kho chnh .....................................................................................152 nhng cng trnh ca tc gi cng cc cng tc vin cng b.....................155

    C S V PHNG PHP SINH HC PHN T .....................................................162

    3

  • Li ni u Sinh hc phn t hin i ang pht trin mnh v tr thnh nng ct ca Cng ngh sinh hc. Vit Nam, thnh tu nghin cu Sinh hc phn t v p dng k thut phn t trong nghin cu Khoa hc s sngv Cng ngh sinh hc c nhiu ng gg trong vic chm sc, bo v sc kho con ngi; trong nh gi ti nguyn sinh vt; trong chn ging v sn xut nng lm ng nghip. Sinh hc phn t l mn Sinh hc hin i, c ging dy nhiu trng i hc v ang c nhng ng gp nht nh trong o to lp ngi c tri thc v Cng ngh sinh hc gp phn vo s nghip Cng nghip ho, hin i ho C s v phng php Sinh hc phn t c bin son t nhiu ti liu, bi ging v cng trnh nghim cu mi v Sinh hc phn t hin i ca cc tc gi trong v ngoi nc, nhm cung cp nhng kin thc c bn v nguyn l v ng dng ca Sinh hc phn t, lm ti liu cho nghin cu, ging dy v hc tp mn ny trng i hc. Cun C s v phng php Sinh hc phn t c cu trc bi 9 chng: Chng 1. H gene

    Chng 2. c im cu trc gene ca sinh vt Prokaryot v Eukryot Chng 3. Mi lin h gia ADN, ARN, Protein

    Chng 4. Enzyme s dng trong k thut sinh hc phn t Chng 5. Xc nh mi lin quan gia protein v c tnh sinh vt Chng 6. K thut sinh hc phn t trong tch h gene sinh vt Chng 7. Phn ng chui polimerase (PCR) Chng 8. T bo ch v Vector Chng 9. Phn lp gene, tch dng phn t v biu hin gene.

    Tc gi trn trng cm n PGS.TS. Nguyn Trng Lng, TS. Lng Th Hng Vn c v gp cho bn tho, xin cm n nhng ng gp ca Hi ng nghim thu nh gi gio trnh ca trng i hc S phm - i hc Thi Nguyn v cm n cc kin ng gp qu bu ca ng o cc nh khoa hc. Trong qu trnh bin son chc chn c nhng sai st, tc gi rt mong nhn c nhng gp ca bn c. Mi ng gp xin gi v Khoa Sinh - K thut Nng nghip trng i hc S phm - i hc Thi Nguyn.

    Tc gi

    4

  • Chng 1: H GENE Tm tt: S ra i ca sinh hc phn t c nh du bng thi im m Oatsn v Cric (1953) pht hin ra cu trc ADN. Tri qua hn 40 nm (1953 - 2000) sinh hc phn t t c nhng thnh tu v i m nh cao ca s pht trin ny l nhng khm ph bn cht sinh hc ca s sng cp phn t v xy dng cc k thut sinh hc phn t ng dng vo thc tin. Geneomics v Proteomics l vn ang c c bit quan tm hin nay m c s ca cc lnh vc ny l nhng pht hin v cu trc v chc nng ca axit nucleic, v c im ca genome nhn, genome ti th, genome lp th. Nhng im khc nhau v cu trc v chc nng ca cc h gene cho php ng dng vo thc t chn ging v nghin cu ngi. Cng vi cu trc ADN v ARN cn c c im ca qu trnh ti bn ADN v phin m cng c quan tm, v n l c s ca nhng k thut sinh hc phn t - cc thao tc ADN v ARN. Ni dung c bn ca chng cp n nhng c s ca sinh hc phn t. Ni dung ca chng gm 4 vn c bn: (1). Khi nim h gene; (2). Axit nucleic; (3). ADN v ti bn ADN, (4). ARN v c ch phn m.

    1. KHI NIM H GENE (GENOME) Qu trnh sinh trng v pht trin ca sinh vt tri qua rt nhiu giai on v tt c qu trnh u ph thuc vo s iu khin ca cc gene. Cu trc, chc nng ca t bo c quyt nh trc tip bi protein, l sn phm cui cng ca s biu hin gene. Qu trnh th hin hot ng ca gene qua protein, b nh hng rt ln bi cc yu t ngoi cnh nh nh sng, ch dinh dng, s cng sinh v s tng tc ca cc gene trong h gene ca t bo. Nh vy, gia cc thnh phn ca h gene trong t bo sng c s tng tc vi nhau v c mi quan h vi cc yu t ca mi trng. Trong t bo, bn cnh genome trong nhn, h thng di truyn cn phn b trong lc lp, ti th v plasmid (vi khun). Cc c quan t ny cng tham gia vo qu trnh sinh tng hp protein ca t bo. T bo l n v cu trc v chc nng c bn ca cc c th sng, t bo cn l n v ca s di truyn. H gene l ton h cc gene trong t bo ca c th sinh vt, h gene ha ton b thng tin di truyn c trng cho tng loi, cho tng c th trong loi. sinh vt Prokaryot h gene gm ton b cc gene trong t bo; cn Eukaryot

    5

  • h gene gm ton b cc gene trong t bo n bi (n). T bo n bi c mt h gene, cc sinh vt hoc t bo lng bi c hai h gene, sinh vt a bi c nhiu h gene. H gene ca sinh vt Eukaryot bao gm h gene nhn (genome nhn), h gene ti th (genome ti th), h gene lc lp (genome lc lp). H gene Prokaryot ch c

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