IV.3.4.

c im v chc nng ca RNA c im

ARN l acid nucleic c thnh

phn ng riboza ARN cu to t 1 chui polyribonucleotide nhng cng c nhng on to lin kt b sung gia hai phn khc nhau ca chui, trong , A lin kt vi U thay cho T C nhiu loi ARN c cu to chc nng khc nhau,song c 3 loi ch yu mARN ,tARN ,rARN.

Chc nng ARN

m-ARN
mARN c tng hp trong dch nhn t ADN. mARN c i sng rt ngn ARNm c ti to rt nhanh v n ch tn ti trong thi gian ca mt thng tin Mt ARNm c th c c nhiu ln nu tin hnh dch m trn polyribosome. m- ARN mang thng tin di truyn tng hp nn 1 loi protein

t -ARN
tARN l mt mch polynucleotit c t 80 n 100 ribonucleotit t xon nh hnh chc ba nn mt s on c nguyn tc b sung A-U; G-C C nhng on khng c nguyn tc b sung, nhng on ny to thnh cc thy trn Mt trong cc thy trn mang b ba i m. Mt si mt ca si ARN 3' gn vi mt acid amin v u mt t do 5' tARN ch vn chuyn mt loi acid amin ng vi b ba i m m mang.Cc acid amin s c vn chuyn n riboxom tng hp protein

r- ARN
ARNr c tng hp trong nhn con v ngay sau lin kt vi protein to nn cc phn t ribonucleoprotein l cc tin ribosome. Qua qu trnh trng thnh, cc ribonucleoprotein ny chuyn t nhn con ra t bo cht v to thnh ribosome

Tng hp RNA in vitro (in vitro transcription)

1. Set up reaction: Mix 1 mL of linearized DNA template pBSSK- ERD (X-E) (0.1-0.5 mg), 7.8 mL of dH2O, 5 mL of Stratagene 5X transcription buffer, 1 mL of 750mM DTT, 1 mL of Rnasin (RNase inhibitor; 15U/ml), 1 mL of 10mM ATP (pH 7), 1 mL of 10mM CTP (pH 7), 1 mL of 10mM GTP (pH 7), 1 mL of 1 mM UTP (pH 7), 5 mL of (a-32P )-UTP (400-800 Ci/mmol; 10 mCi/mL) (final conc. 4-8mM), 0.2 mL T7 or T3 or SP6 RNA polymerase (50 U/mL, Stratagene). Total volume: 25 mL 2. Mix, spin, and incubate at 37oC for 30 minutes 3. Add 1 mL of DNase 1 (free from Rnase -10 U/mL) 4. Mix, spin, and incubate at 37oC for 15 minutes 5. End the reaction by adding: 7 mL of 10X STE (0.2 M Tris-HCl, pH 7.5; 1 M NaCl; 0.1 M EDTA) and 38 mL of dH2O6. Mix by pipeting and spot 1 mL on paper disk inside a scintillation bottle7. Purify the rest of the probe through a gel filtration PUSH COLUMN (Stratagene)