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한우에 감염된 Theileria sergenti merozoite의 순수분리와 genomic DNA probe에 관한 연구
채준석,이주묵,권오덕,채건상 全北大學校 基礎科學硏究所 1994 基礎科學 Vol.17 No.-
To make the genomic DNA probe of Theileria sergenti, the merozoites were purified from bovine crythrocytes. The infected erythrocytes were lysed by Aeromonas hydrophila(Ah-1) hemolysin, and the parasites were isolated by ultracentrifugation on a Percoll discontinuous density gradient. For construction of a T sergenti genomic DNA library, T sergenti DNA was digested with PstI and the fragments were ligated into the PstI site of pUC19 before transformation of Escherichia coli JM83. Out of thousands of transformants obtained by transformation of E coli JM83 with the genomic library, three plasmids were chosen. The sizes of the inserted DNAs were 2.9kb(2.4kb and 0.5kb) in pKTS1, 4.3kb in pKTS2 and 1.5kb in pKTS3, respectively. The DNA fragments used as probe KTS1(2.4kb), KTS2(4.3kb) and KTS3(1.5kb) were labeled digoxigenin-11-dUTP for the Southern hybridization. In Southern hybridization, all of the probes(KTS1, KTS2 and KTS3) reacted specifically to T sergenti DNA, but not to bovine leucocyte DNA. In order to find out the sensitivities of the digoxigenin-11-dUTP-labeled KTS1 and KTS3 as the probes, purified merozoite DNA and bovine DNA(control) were checked by dot blot hybridization with the probes. Both of the probes, KTS1 and KTS3, detected as minimum amount of 975pg of the T sergenti DNA, but not bovine DNA even to 500ng.
Si Bulk Micromachining을 위한 Wafer Rolling Etching 및 그 특성
김건년,이보나,박효덕,신상모,공경준,장동근,김병철,권혁채,이봉희 경북대학교 센서기술연구소 1998 센서技術學術大會論文集 Vol.9 No.1
A wafer rolling etching system for the silicon bulk micromachining has been designed and fabricated. The silicon diaphragms were anisotropically etched in a 24.5 weight percent KOH solution. Compared to the conventional KOH etching systems, pyramidal hillocks, and wave-shaped structures on the etched surfaces were greatly reduced by using this system. After etching for time of 438 minutes, the average etched depth and the etch-rate were measured to be 537μm and 1.22μm/min, respectively. The average etching uniformity of etching depth was 0.87% in 5-inch wafer. Our results showed that the wafer rolling method enhanced etch uniformity and etch rate.
Chae, Keon-Sang,Katsuji Murakawa,Kousaku Okubo,Kenichi Mastsubara 全北大學校 基礎科學硏究所 1994 基礎科學 Vol.17 No.-
For the PCR-based chromosomal assignment of very short cDNA fragments specifically designed primers are required. We tested primers with very short core sequences that are identical or complementary to known cDNA sequences, with or without tails at the 5' ends. The lower limit of the core length for PCR using human chromosome templates was 14 nucleotides (nt) when they have tails. The minimal length of the tail was 2 nt when it was attached to the 5' end of a 14-nt core. In the absence of a tail. 15 nt are needed for the core to act properly. The overall size of the short cDNA fragments that could be assigned was further reduced by using a pair of primers that overlap at the 3' ends. The limits of the free energy of overlap were about -1.9 kcal mol at 45 C, -2.9 ㎉/mol at 50 C and -4.5 ㎉/mol at 55 C. A combination of these features in a primer pair allowed cDNA fragments as short as 30 nt to be assigned.
Chromosomal Assignment of Short cDNA Sequences by PCR Using Overlapping and Tailed Short Primers
CHAE, Keon-Sang,MURAKAWA, Katsuji,OKUBO, Kousaku,MATSUBARA, Kenichi 全北大學校 基礎科學硏究所 1994 基礎科學 Vol.17 No.-
Overlapping primers and tailed short primers are effective agents for mapping very short cDNA sequences. By using such primers, human cDNAs as short as 32 nucleotides in length can produce PCR bands. Using these and other primers of ordinary size, 44 cDNAs were assigned to chromosomes, of which 24 were assigned to single chromosomes, and 2 were assigned to two chromosomes and two were assigned to three chromosomes, respectively. Among the 24 cDNAs, all of which matched GenBank entries, 6 cDNAs were observed to map to the same chromosomes as reported previously.
Proteus vulgaris로부터 N4 파아지에 대해 내성을 갖게 하는 유전자의 분리
채건상 전북대학교 유전공학연구소 1988 遺傳工學硏究所報 Vol.1 No.1
대장균에 감염하는 파아지 중의 하나인 N4가 대장균에 감염하지 못하도록 해주는 rtn(resistant to N4)유전자를 Proteus vulgaris로부터 대장균에 분리하였다. 이 rtn 유전자를 갖고 있는 재조합 플라스미드인 pRMG101을 갖고 있는 대장균은 전부 N4의 감염을 저해하였으나, 다른 파아지인 T4, T7, Ø80, lambda가 감염하는 데에는 아무 영향을 주지 못하였다. 이 유전자의 전체 염기 서열을 밝혀본 결과, promoter 부분과 두 개의 단백질을 만들 수 있는 open reading frame(rtn A와 rtn B)이 있다는 것을 알아내었는데 이 rtn 유전자의 promoter는 대장균내에서도 자신의 기능을 발휘하는 것이 확산되었다.
rtn(resistant to N4) 유전자의 정체 : 새로운 기능의 유전자 a set of Novel Genes with a new Function
채건상 全北大學校 基礎科學硏究所 1988 基礎科學 Vol.11 No.1
Proteus vulgaris로부터 분리된 rtn (resistant to N4)유전자를 갖고 있는 대장균에 N4는 감염하지 못하였으나 lambda, Φ80, T4, T7등의 다른 파아지는 감염할 수 있었다. rtn유전자를 갖고 있는 재조합 플라스미드 pRMG101은 PvuⅠ과 PvuⅡ 제한효소에 의해 완전히 절단되었다. 또한 이 재조합 플라스미드를 갖고 있는 대장균을 키워 제 2형의 제한-변형효소의 활성을 찾아보았으나 그런 활성은 나타나지 않았다. 이러한 결과들은 rtn 유전자가 지금까지 알려진 제 2형의 제한-변형효소의 유전자는 아니라는 것을 보여준다. 뿐만아니라 제 1형 제한-변형효소의 특성과 rtn 유전자와 비교한 결과와 rtn 유전자를 갖고 있는 대장균을 N4의 DNA만으로 형질전환시켰을 때 정상적인 N4 파아지가 나올 수 있음을 확인한 결과로 미루어 rtn 유전자가 대장균에의 N4 파아지 감염을 저해하는 기작은 P.vulgaris의 제한-변형효소에 의한 것 또는 세포내에서 일어나는 것은 아니고 세포밖에서 N4가 감염하는 과정을 저해하는 것으로 생각된다.