지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 1995.8
- 수록면
- 209 - 220 (12page)
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The nuclear matrix was isolated from Misgurnus mizolepis liver nuclei by low salt extraction and restriction enzyme treatment. The structure was digested with proteinase K. After centrifugation, matrix attachment regions (MARs) were obtained by RNase treatment and phenol-chloroform extraction. The result leads to the appearance of smeared bands in the range of about 0.3-15 kb.
pURY19 vector was constructed by inserting 2.13 kb Eco47 Ⅲ fragment of the yeast uracil 3 gene into the unique Ssp Ⅰ site of pUC19 plasmid vector as a selection marker. This vector is unable to be maintained in Saccharomyces cerevisiae by itself since it cannot replicate as an extrachromosomal element. Using this system, we attempted cloning the ARS (autonomously replicating sequence) from M. mizolepis to develop an efficient expression vector for the transgenic fish. pURY19N_(1-62) were constructed by inserting MARs in pURY19 plasmid vector and transformation of E. coli DH5α.
Replication origins (ARS) of M. mizolepis were isolated, which enabled the vector to replicate autonomously in S. cerevisiae. The cloned DNA fragments were sequenced by Sanger's dideoxy-chain termination method. All clones were AT-rich. pURY19N_6, one of the clones, expecially contained ARS consensus sequence, Topoisomerase Ⅱ consensus, near A-box and T-box.
pURY19 vector was constructed by inserting 2.13 kb Eco47 Ⅲ fragment of the yeast uracil 3 gene into the unique Ssp Ⅰ site of pUC19 plasmid vector as a selection marker. This vector is unable to be maintained in Saccharomyces cerevisiae by itself since it cannot replicate as an extrachromosomal element. Using this system, we attempted cloning the ARS (autonomously replicating sequence) from M. mizolepis to develop an efficient expression vector for the transgenic fish. pURY19N_(1-62) were constructed by inserting MARs in pURY19 plasmid vector and transformation of E. coli DH5α.
Replication origins (ARS) of M. mizolepis were isolated, which enabled the vector to replicate autonomously in S. cerevisiae. The cloned DNA fragments were sequenced by Sanger's dideoxy-chain termination method. All clones were AT-rich. pURY19N_6, one of the clones, expecially contained ARS consensus sequence, Topoisomerase Ⅱ consensus, near A-box and T-box.
목차
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
요약
REFERENCES
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UCI(KEPA) : I410-ECN-0101-2009-529-015332279