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유전자 재조합 대장균으로부터 인간 Interferon - α2 의 정제
이진규,강인철,박성희,정광희,문홍모 ( Jin Kyu Lee,In Chul Kang,Sung Hee Park,Kwang Hoe Chung,Hong Mo Moon ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.1
Recombinant human IFN-α2 was purified from E. coli by methods involving sonication, extraction with 8 M Guanidine-HCI, dilution, CuSO₄ fractionation, copper-chelating column chromatography, S-Sepharose column chromatography. Specific activity of purified IFN-α2 was 1.6 × 10^9 IU/㎎ protein and the degree of purification was 94 fold from the starting material. Purified IFN-α2 was formed a single band on SDS-PAGE under reducing condition and also on non-reducing condition. Its molecular weight was estimated to be 18,000 dalton and the isoelectric point of purified IFN-α2 was 6.0. Purity of the recombinant IFN-α2 was better than 99% by densitometric analysis of SDS-PAGE.
cDNA Cloning and Expression of a Human Interferon-$\gamma$ in E. coli
고형곤,현형환,유무영,권병세,주종광,문홍모,김광수,Koh, Hyung-Kon,Hyun, Hyung-Hwan,Yoo, Moo-Young,Kwon, Byung-S.,Jue, Chong-K.,Moon, Hong-Mo,Kim, Kwang-Soo 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.3
인간의 peripheralblood lymphocyte에서 분리된 poly (A)$^+mRNA$를 사용하여 ${\lambda}gt$ 10 cDNA library를 제조하였으며, 화학적으로 합성한 oligonucleotide를 이용하여, 이 cDNA library로부터 IFN-$\gamma$ 유전자를 갖는 DNA을 분리했다. 분리된 clone 가운데 하나인 pBK04는 IFN-$\gamma$의 전체 염기서열을 갖고 있었다. IFN-$\gamma$를 E. coli내에서 발현시키기 위해 lpp promotor를 함유하고 있 는 pINIIIA3 vector를 사용하여 IFN-$\gamma$를 발현시킬 수 있는 재조합 plasmid ($pKS{\gamma}3B$)를 제조하였다. $pKS{\gamma}3B$로써 형질 변환된 bacterial cell을 antiviral activity에 의해 역가 분석을 해본 결과, 높은 정도로 IFN-$\gamma$를 발현시켰으며 이 IFN-$\gamma$는 천연의 IFN-$\gamma$에 대하여 특이성이 있는 monoclonal antibody에 의해서도 인지되었다. High copy number를 갖는 expression vector를 제조 (pHK-3) 하였으며, 이 high copy number vector를 사용하므로써 IFN-$\gamma$의 생성이 증가되는 사실을 확인하였다. A ${\lambda}gt10$ cDNA library constructed with poly $A^+$ mRNA from human peripheral blood lymphocytes was screened by a synthetic oligonucleotide having IFN-$\gamma$ sequence. One of the clones (pBK04) which hybridized to the oligomer was confirmed to have the whole IFN-$\gamma$ coding sequence. The eDNA insert was used to express IFN-$\gamma$ in E. coli under the control of lpp promoter employing pINIIIA3 vector. Cells harboring the recombinant plasmid ($pKS{\gamma}3B$) expressed high level of IFN-$\gamma$ assayed by antiviral activity, The molecule was also recognized by monoclonal antibody specific to a natural IFN-$\gamma$. An expression vector with high copy number (pHK-3) was constructed and was successfully used for the improvement of the expression of IFN-$\gamma$.
혈액응고 제8인자 유전자 내부의 BclI, Xbal RFLP 와 St14 VNTR 에 나타나는 다형인자의 빈도추정과 A 형 혈우병 8가계의 연관 분석
문홍모,김장성,오달균,강신혜 한국유전학회 1993 Genes & Genomics Vol.15 No.4
With the acid of PCR, we used two intragenic factor VIII gene RFLP markers(BclI-intron 18 and XbaI-intron 22) and one extragenic marker(St 14 VNTR) to estimate allele frequencies of these markers from 100 Korean individuals and to analyze eight Korean hemophilia A families. Allele frequencies for the presence of the enzyme sites and heterozygosity expectations were 86 and 24% for BclI-intron 18 RFLP marker, and 62% and 47% for XbaI-intron 22 RFLP marker, respectively. When BclI-intron 18 was homozygous, 31.5% of women was predicted to be heterozygous for XbaI-intron 22. Number of repeats in St14 VNTR loci from Korean males differed from those previously reported for Caucasian males. 68% of Korean women was estimated to be heterozygous for St14 VNTR marker. Analysis of hemophilia A families with BclI-RFLP and St14 VNTR proved to be informative for six out of eight families tested.
Squalene Adjuvant를 이용한 단백질 항원의 세포성 면역반응 유도
도현주,김성열,안종성,하영주,이승찬,오재택,장명호,정홍석,문홍모,박해준 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.3
To investigate the role of adjuvant for cytotoxic T lymphocytes (CTL) induction, squalene-based adjuvant was studied its capability eliciting to the induction of cellular immunity as well as humoral immunity to exogeneous proteins. Ovalbumin (OVA) was used as model proteins. It was demonstrated that antigen formulation consisted of metabolizable oil squalene mixed with Tween 80 and pluronic L121 (S/TJ121) could induced the antigen specific CTL responses and antibodies irrespective of immunization routes in mice. We also demonstrated that this antigen formulation was a inducer of CD8+, major histocompatibility complex(MHC) class I-restricted, and antigen-specific C;1'Ls. These data suggest that the squalene-based emulsion system is a potent adjuvant inducing cellular immunity as well as humoral immunity.
대장균에서의 인간 Interferon - γ의 cDNA 크로닝 및 발현
김광수,현형환,유무영,고형곤,권병세,주종광,문홍모 생화학분자생물학회 1991 BMB Reports Vol.19 No.3
A λ gt10 cDNA library constructed with poly A^+ mRNA from human peripheral blood lymphocytes was screened by a synthetic oligonucleotide having IFN-γ sequence. One of the clones (pBK04) which hybridized to the oligomer was confirmed to have the whole IFN-γ coding sequence. The cDNA insert was used to express IFN-γ in E. coli under the control of lpp promoter employing pINTIIIA3 vector. Ce115 harboring the recombinant plasmid (pKS γ 3B) expressed high level of IFN-γ assayed by antiviral activity. The molecule was also recognized by monoclonal antibody specific to a natural IFN-γ. An expression vector with high copy number (pHK-3) was constructed and was successfully used for the improvement of the expression of IFN-γ