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조명선,정태진,김남덕,정재성,정재훈,이영훈,Cho, Myung-Sun,Cheong, Tae-Chin,Kim, Nam-Deok,Jung, Jae-Sung,Chung, Jae-Hoon,Lee, Young-Hoon 생화학분자생물학회 1992 한국생화학회지 Vol.25 No.6
정상적인 온도에서 생장하고 있는 박테리아 배양액을 저온으로 옮겨 주었을 때 생성이 증가되는 단백질의 양상을 1차원 및 2차원 전기영동에 의해 조사하였다. 이러한 저온 충격단백질들의 생성은 충격을 받은 온도에 따라 다소 다르게 나타났으며 일단 생성된 저온 충격단백질들은 $37^{\circ}C$ 로 다시 온도를 옮겼을 때에도 안정한 것으로 관찰되었다. 저온 충격단백질의 생성기작을 이해하기 위해 저온 충격단백질로 알려진 RecA의 유전자를 pUC19에 클로닝한 후 재조합 플라스미드를 함유하는 군주들에서 저온 충격에 의해 RecA 단백질의 생성이 증가됨을 분자수준에서 증명하였다. The cold shock proteins induced by temperature shift from $37^{\circ}C$ to $10^{\circ}C$ were assayed by the two-dimensional polyacrylamide gel electrophoresis after the proteins were pulse-labeled with $^{35}S$-methionine. The cold shock proteins were quite stable even if the temperature was returned to $37^{\circ}C$. The cloned recA gene was used to understand the molecular mechanism of the cold shock response. Synthesis of RecA protein from the recA gene harbored on the multicopy plasmid was increased upon cold shock, suggesting that the cold shock response involves the regulation by unlimited cellular factors such as RNA polymerase.
조명선,정태진,김남덕,정재성,정재훈,이영훈 ( Myung Sun Cho,Tae Chin Cheong,Nam Deok Kim,Jae Sung Jung,Jae Hoon Chung,Young Hoon Lee ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.6
The cold shock proteins induced by temperature shift from 37℃ to 10℃ were assayed by the two-dimensional polyacrylamide gel electrophoresis after the proteins were pulse-labeled with ^(35)S-methionine. The cold shock proteins were quite stable even if the temperature was returned to 37℃. The cloned recA gene was used to understand the molecular mechanism of the cold shock response. Synthesis of RecA protein from the recA gene harbored on the multicopy plasmid was increased upon cold shock, suggesting that the cold shock response involves the regulation by unlimited cellular factors such as RNA polymerase.
조명선,정태진,정재성,이영훈,조철오,정재훈 ( Myung Sun Cho,Tae Chin Cheong,Jae Sung Jung,Young Hoon Lee,Cheol O Joe,Jae Hoon Chung ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.8
Temperature downshift from 37℃ to 10℃ or 18℃ induces over 10 cold shock proteins in Escherichia coli. We have reported that the synthesis of RecA, one of the cold shock proteins, was increased two-fold upon cold shock when it was expressed in multicopy plasmid. In an attempt to understand the molecular mechanism of cold shock response, the expression of the recA gene was examined at transcriptional level in this study. Northern blotting and chloramphenicol acetyl transferase activity from the recA-CAT fusion plasmid revealed that the recA transcription was increased 2∼3 fold by cold shock, indicating that the cold shock response is at the level of transcription. The gel mobility shift experiment suggested that cold shock might induce the transcription by releasing the regulatory factor(s) from the recA promoter region.
조명선,정태진,정재성,이영훈,조철오,정재훈,Cho, Myung-Sun,Cheong, Tae-Chin,Jung, Jae-Sung,Lee, Young-Hoon,Joe, Cheol-O,Chung, Jae-Hoon 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.8
저온 충격단백질의 생성기작을 분자수준에서 이해하고자 저온 충격단백질의 하나로 알려진 RecA를 선택하여 이것의 저온 충격에 의한 생성 증가기작을 연구하였다. Northern 브럿에 의한 전사체 양의 비교 및 융합된 CAT(크로람페니콜 아세틸기 전달효소) 유전자의 발현을 통하여 저온 충격에 의해 recA 유전자의 발현이 2배 증가되는 것이 관찰되었다. 이러한 결과로 미루어 저온 충격 대응기작이 전사단계에서 이루어짐을 알수 있었다. 또한 저온 충격에 관여하는 조절인자의 존재여부를 조사하기 위하여 recA 유전자의 촉진유전자 부위에 결합하는 단백질에 대한 실험결과 recA 유전자의 촉진유전자에 조절인자가 결함하는 것이 저온 충격에 의해 오히려 억제되는 것으로 나타났다. Temperature downshift from $37^{\circ}C$ to $10^{\circ}C$ or $18^{\circ}C$ induces over 10 cold shock proteins in Escherichia coli. We have reported that the synthesis of RecA, one of the cold shock proteins, was increased two-fold upon cold shock when it was expressed in multicopy plasmid. In an attempt to understand the molecular mechanism of cold shock response, the expression of the recA gene was examined at transcriptional level in this study. Northern blotting and chloramphenicol acetyl transferase activity from the recA-CAT fusion plasmid revealed that the recA transcription was increased 2~3 fold by cold shock, indicating that the cold shock response is at the level of transcription. The gel mobility shift experiment suggested that cold shock might induce the transcription by releasing the regulatory factor(s) from the recA promoter region.
Serratia marcescens 의 R plasmid 로부터 gentamicin 저항성을 갖는 유전자의 cloning
신창호,김인규,정재훈,최영길,연승우,조명선,정재성 한국유전학회 1991 Genes & Genomics Vol.13 No.3
Gentamicin resistance gene residing in Serratia marcescens R plasmid pIS199G2 was cloned after it was transferred to Escherichia coli ML1410 by conjugation. Partial digests of pIS199G2 with Sau3A1 were cloned into the BamHI site of pUC9. One plasmid pSY1 carried 1.5kb insert and showed gentamicin resistant phenotype. A polypeptide product of 31kd was identified from pSY1 in minicell expression system. During the cloning procedure a plasmid narboring a replication origin in R plasmid, ampicillin resistance gene and gentamicin resistance gene was obtained. The plasmid, pSW1, also produced the polypeptide of the same size by the minicell expression system. The restriction enzyme mapping of pSY1 and pSW1 showed homology to other aacC2 type of gentamicin resistance gene and suggests that the two plasmids contain aacC2 gene family.