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형질전환체 담배에서 사람 파필로마 바이러스 HPV - 18 E6 유전자의 발현
전재필,현미아,김달웅,진승원,박순희,김현수,유왕돈,손우익 ( Jaepil Jeon,Meeah Hyun,Dalung Kim,Seungwon Jin,Soonhee Park,Hyunsoo Kim,Wangdon Yoo,Uik Sohn ) 생화학분자생물학회 1994 BMB Reports Vol.27 No.5
To develop a HPV-18 oral vaccine, HPV-18 E6 gene was cloned into an expression vector, under the control of CaMV 35S dual promoter and 5`UT region of TEV. The constructed vector (pUS-E6) was used to transform tobacco plants by the Agrobacterium-mediated leaf disc method. The transformants were then regenerated to whole transgenic plants. The HPV-18 E6 gene was found to be expressed in the transgenic tobacco plants as determined by PCR, RT-PCR, Northern analysis and Western analysis. These results show a possibility that HPV oral vaccine may be developed by expressing the HPV proteins in tomato or other vegetables.
Ribosomal DNA의 internal transcribed spacer 염기서열과 RAPD 비교에 의한 작약류의 유연관계 분석
Jeong Hye Kim(金廷憓,),Uik Sohn(孫宇翼),Hyun Cheol Lee(李鉉喆),Jae Chui Kim(金在喆),So Deuk Park(朴小得) 한국육종학회 1997 한국육종학회지 Vol.29 No.3
To analyse the genetic relationship among 14 peony accessions, ITS sequencing of rDNA and RAPD were exploited. We amplified 653bp regions including ITS(Ⅰ), 5.8S rDNA and ITS (Ⅱ). The amplified regions were sequenced and compared with GenBank database. The sequences of 5.8S rDNA region were identical among peony species and sub-species. Nucleotide differences at 16 different sites in ITS (Ⅰ) region were observed. In ITS (Ⅱ) region, 1 base pair deletions at one nucleotide site and nucleotide differences at 13 different sites were observed among sequences of 14 peony accessions and GenBank sequences. And at certain nucleotide sites that show nucleotide differences, two different nucleotides were observed in one nucleotide position. Results of ITS sequence analysis were consistent to the classification of peony species based on morphological characteristics. These results suggest that we can distinguish peony species by ITS sequence comparison. We also utilized RAPD analysis in order to study the phylogenetic relationship among peonies. Initially we screened 30 RAPD primers using genomic DNA and selected 11 primers showing distinct band patterns. Ninety eight reproducible bands were generated by 11 primers and the band patterns were analysed to generate dendrogram showing genetic diversity by UPGMA method. As a result, 14 peony accessions were classified into group A, B, C and D. Group D was subdivided into D-1, D-2 and D-3. This study shows that RAPD is a rapid and efficient tool for phylogenetic grouping of peony species and subspecies.
김성환(Sung Whan Kim),김삼묘(Sam Myo Kim),이창묵(Chang Muk Lee),손우익(Uik Sohn) 한국정보과학회 1998 한국정보과학회 학술발표논문집 Vol.25 No.1A
본 논문은 Leonard Adleman의 Hamiltonian 경로 탐색을 위한 DNA 계산방법이 보다 다양한 문제에 효율적으로 적응할 수 있는 방안을 실험을 통하여 개발하는데 그 목적이 있다. 특히 이 논문은 합성해야 할 DNA 서열의 선택, 그래프의 특성 및 해밀토니안 경로의 다양성에 따라 새로운 실험 프로토콜을 개발하였다. 무작위로 만든 서열에서 선택한 서열을 합성하여 ligate 하고 증폭(PCR)하여 해를 구하는 최적 조건은 Adleman이 보고한 것과는 현저히 다름을 알 수 있었으며, 그래프가 많은 짧은 순환경로는, 우려했던 것과는 달리 실험에 장애가 되지 않았다. Adleman이 제시한 방법은 해밀토니안 경로의 존재 여부 만을 알려준다. 본 논문은 그래프에 경로가 하나 이상 있을 때 그 경로들을 개별적으로 선별해 낼 수 있는 방법을 실험을 통하여 보여준다.
자궁상피내암종 및 침윤성 편평상피암에서의 In Situ Hybridization 에 의한 Human Papillomavirus DNA 의 검색
손우익,진승원,이상숙,김기권,정재홍 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
The authors examined 9 condylomas, 26 cervical intraepithelial neoplasms(CIN) and 22 invasive squamous cell carcinomas for the presence of human papillomavirus(HPV) DNA sequences by DNA-DNA in situ hybridization. In situ hybridization revealed target HPV DNA sequences mostly in the nuclei of the superficial cells from epithelium which contained either maturation or koilocytotic atypias. With the use of biotinylated HPV DNA probes 6/11, 16/18 and 31/33/35, 42 of the 57(73.7%) were positive with any HPV DNA mixed probes, 13 of which were positive with HPV-6/11, 23 with HPV-16/18, 32 with HPV-31/33/35 and 18 with two or more mixed probes. HPV-31/33/35 was the most prevalent in CIN and invasive squamous cell carcinomas, followed by HPV-16/18. The incidence of HPV DNA increased from 66.7% to 86.496 with increasing severity of the lesions from condylomas to invasive squamous cell carcinomas. Flat condyloma was most frequently accompanied by CIN.
손우익,이상숙,김기권,박준식,진승원 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
Verrucous carcinoma of the larynx is a distinct and uncommon variant of well-differentiated squamous cellcarcinoma. The authors hybridized in situ a case of laryngeal verrucous carcinomma with DNA probes to human papillomavirus (HPV) 6, 11, 16, 18, 31, 33 and 35. The DNAs from the verrucous carcinoma of larynx hybridized with mixed DNA probes HPV-16/18 and 31/33/35. In addition, there was an evidence of HPV infection based on hybridization with mixed DNA probe HPV-6/11 in the adjacent papilloma tissue. By in situ DNA hybridization techniques, we clearly demonstrated human papillomavirus (HPV-16/18, and 31/33/35) -related sequences in this neoplasm. These findings suggest the role of HPV-6/11 in the development of laryngeal papilloma and HPV-16/18 and 31/33/35 probably on the progression to verrucous carcinoma.
자궁경부의 상피내암 조직내 In Situ Hybridization 에 의한 Human Papillomavirus DNA 의 검색 및 Typing
손우익,이상숙,이태성,김기권,정재홍,김택훈,진승원 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
The authors examined 12 condylomas, 55 cervital intraepithelial neoplasms(CIN) for the presence of human papillomavirus(HPV) DNA sequences by DNA-DNA in situ hybridization and reviewed the epithelial distribution and incidence of HPV DNA sequences in different cervical lesions. In situ hybridization to HPV DNA probes revealed target HPV DNA sequences mostly in the nuclei of the superficial cells from epithelium which contained either maturation of koilocytotic atypias, although in several cases hybridizing sequences were detected in the entire thickness of epithelium, including basal layer. The distribution of HPV was mostly focal and multicentric. With the use of biotinylated HPV DNA probes 6/11, 16/18 and 31/33/35, 41 of 55(74.5%) cases of CIN was positive with any HPV DNA probes, 20 of which were positive with HPV 6/11, 28 with HPV 16/18, 36 with HPV 31/33/35 and 27 with two or more mixed probes. HPV 31/33/35 was the most prevalent in CIN grade Ⅲ, followed by HPV 16/18, although there was no difference in the frequency of HPV types among condylomas, CIN grade Ⅰ and CIN grade Ⅱ. Cases of CIN under the age of 40 revealed higher HPV-positive rate and more abundant HPV DNA than those with age 40 or more. Mixed HPV infection, positive with 2 or more kinds of HPV DNA probes was found about half of the cases studied. Among type of condylomas, flat condyloma was the most frequently accompanied by CIN.
HPV (Human papillomavirus) 양성인 자궁경부암에서의 세포성 면역 반응
손우익,이상숙,이태성,진승원 경북대학교 유전공학연구소 1992 遺傳工學硏究所報 Vol.7 No.1
To study the human immune response to HPV, three monoclonal antibodies against T cells(UCHL), B cells(L-26), and macrophages (CD68) were used in an immunohistochemical study of HPV-infected cervical cancer tissues. Most infilterlating leucocytes were found to be predominantly T cells and macrophages, with few B cells in all tissues examined. These results support the hyopothesis that cell-mediated immunity is related to the HPV-induced disease.