DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | Kim, Hyung-Man | - |
dc.contributor.advisor | 김형만 | - |
dc.contributor.author | Hong, Soon-Kwang | - |
dc.contributor.author | 홍순광 | - |
dc.date.accessioned | 2011-12-12T08:56:17Z | - |
dc.date.available | 2011-12-12T08:56:17Z | - |
dc.date.issued | 1984 | - |
dc.identifier.uri | http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=64063&flag=dissertation | - |
dc.identifier.uri | http://hdl.handle.net/10203/28183 | - |
dc.description | 학위논문(석사) - 한국과학기술원 : 생물공학과, 1984.2, [ viii, 50 p. ] | - |
dc.description.abstract | Plasmid POTC-10 DNA which is a derivative of PBR322 and contains trp-E gene promoter of $\underline{E}$. $\underline{coli}$ was isolated from E. coli HB101. This plasmid DNA was digested with ClaI restriction enzyme and treated with bacterial alkaline phosphatase to inhibit self-ligation. Poly-d(A:T) duplex (about 1 kilo-base-pair) was purchased from P.L. Biochem.. Because poly-d(A:T) duplex was very heterogeneous in molecular size, it was denatured and renatured. ClaI-linker was labelled with $r-^{32}$p ATP. This labelled ClaI-linker was ligated to poly-d(A:T) duplex with $T_4$-DNA ligase and digested with ClaI restriction enzyme. The ClaI digested POTC-10 plasmid DNA and poly-d(A:T) duplex:ClaI-linker were ligated with $T_4$-DNA ligase. This ligate was used to transform $\underline{E}$. $\underline{coli}$ HB101 competent cell treated with $CaCl_2$ buffer solution. Plasmid DNA was isolated from the each transformant (ampcilline plate) by rapid small scale isolation method. Their size was checked on 1% agarose gel electrophoresis and about 15% of the isolated DNA was appeared to have increased size. These plasmid DNA which showed an increase in length were digested with Hind III restriction enzyme and checked on 1% agarose gel electrophoresis whether they were the dimer of POTC-10 or not. From this, it was certain that they may be not the dimer but the recombinant which were inserted with 1 kilo-base paired fragment. Then the recombinant DNA were digested with ClaI restriction enzyme and the existence of poly-d(A:T) duplex was checked by colony hybridization and by southern blotting hybridization technique. | eng |
dc.language | eng | - |
dc.publisher | 한국과학기술원 | - |
dc.title | Cloning of poly-d(A:T) duplex in E. coli | - |
dc.title.alternative | E. coli 에서의 poly-d(A:T) duplex 의 cloning | - |
dc.type | Thesis(Master) | - |
dc.identifier.CNRN | 64063/325007 | - |
dc.description.department | 한국과학기술원 : 생물공학과, | - |
dc.identifier.uid | 000821363 | - |
dc.contributor.localauthor | Kim, Hyung-Man | - |
dc.contributor.localauthor | 김형만 | - |
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