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박선희,이승기,박종상 ( Sun Hee Park,Seung Ki Lee,Jong Sang Park ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.1
K-ras gene expression in the regenerating liver of the patial hepatectomy was tested by using dot hybridization. The K-ras gene transcription after the patial hepatectomy is amplified and its RNA splicing patern is changed. In normal liver exon 4a is spliced out and only K-ras proteins containing exon 4b are produced. In regenerating liver exon 4b is spliced out and K-ras proteins containing exon 4a might be produced at least 1000% more than in the normal liver.
노지애,박선희,박종상 ( Jee Ae Noh,Sun Hee Park,Jong Sang Park ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.1
Human fibroblast Interferon cDNA of which the signal sequence was elimiated by the partial digestion of the restriction enzyme AluI was inserted into the E. Coli expression vector, pkk223-3 which has tac promoter and transcription terminator region, rrnBT₁T₂ and into PLc2833 which has λpL promoter. The former was transformed to JM105 and JM103 and the latter was transformed to NR69 and M5248. The orientation of the IFN-β toward promoter was ascertained by restriction enzyme cutting. Expression of IFN-βgene of pkk223-3-IFN-βin E. Coli was induced by IPTG and the of PLc2833-IFN-β was induced by heat-shock. The induced E. Coli lysates were analysed by using SDS-PAGE.
Cloning and Sequence Analysis of 3'-Terminal Region of Human Insulin Receptor Gene
이정화,박선희,박종상,Lee, Jung-Hwa,Park, Sun-Hee,Park, Jong-Sang Korean Society for Biochemistry and Molecular Biol 1990 한국생화학회지 Vol.23 No.3
사람 인슐린 수용체 유전자의 3'말단 부분의 5 kb를 클로닝하였다. 건조젤 하이브리디제이션을 이용하여 제한효소인 EcoRI으로 자른 사람 인슐린 수용체 유전자 중 4.5-5.5 kb를 확인하고 ${\lambda}gt$ 10벡터에 삽입시켰다. 이렇게 하여 만든 유전자 라이브러리를 카르복시 말단에 해당하는 합성 NDA로 스크린하여 표지를 내는 plague들을 pBR328에 재클로닝하였다. 부분적인 DNA의 염기서열 결정으로 클로닝한 DNA가 사람 인슐린 수용체임을 확인하였다. ExoIII nuclease로 DNA를 크기순서대로 자르는 방법을 이용하여 전사 말단 부위의 1.3 kb의 유전자 염기서열을 결정하였다. 염기서열 분석결과 인슐린 수용체 유전자는 poly(A)가 붙는 자리 앞쪽의 15 뉴클레오티드에 전통적인 AATAAA 염기가 아닌 AATATA 염기가 존재함을 알았는데 아마도 이것이 유전자 조절에 관여할 것으로 여겨진다. 또한 이 유전자는 mRNA 프로세서에 관여한다고 제안된 G/T 밀집 염기서열을 가지고 있음을 알았다. The 5 kb long 3'-terminal region of genomic DNA of Human Insulin Receptor(IR) gene was cloned. By the dry-gel hybridization, 4.5-5.5 kb long DNA of EcoRI cut human chromosomal DNA was identified and inserted into ${\lambda}gt$ 10 vector. Constructed subgenomic library was screened with synthetic 21 mer probe corresponding to the carboxy terminal. By the partial DNA sequencing about 200 bp in 5 kb genomic DNA, we found the sequence was the same as the published result. We sequenced about 1.3 kb transcription terminal region by the ExoIII deletion method. The result of the sequence analysis showed that the IR gene has AATATA sequence 15 nucleotide upstream of poly(A) site instead of the canonical AATAAA consensus sequence and this may play a role in gene regulation and that in poly(A) downstream region the IR gene has the T-rich, G/T cluster sequence, which was proposed as a element of mRNA processing.
박선희,박종상,노지애 생화학분자생물학회 1994 BMB Reports Vol.17 No.3
Human fibroblast Interferon cDNA of which the signal sequence was elimiated by the partial digestion of the restriction enzyme AluI was inserted into the E. Coli expression vector, pkk223-3 which has tac promoter and transcription terminator region, rrnBT₁T₂ and into PLc2833 which has λpL promoter. The former was transformed to JM105 and JM103 and the latter was transformed to NR69 and M5248. The orientation of the IFN-β toward promoter was ascertained by restriction enzyme cutting. Expression of IFN-βgene of pkk223-3-IFN-βin E. Coli was induced by IPTG and the of PLc2833-IFN-β was induced by heat-shock. The induced E. Coli lysates were analysed by using SDS-PAGE.