지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제24권 제11호
- 발행연도
- 2014.1
- 수록면
- 1,503 - 1,509 (7page)
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초록· 키워드
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With the purpose of facilitating the process of stable strain generation, a shuttle vector for integration of genes via a double recombination event into two ectopic sites on the Sulfolobus acidocaldarius chromosome was constructed. The novel chromosomal integration and expression vector pINEX contains a pyrE gene from S. solfataricus P2 (pyrEsso) as an auxotrophic selection marker, a multiple cloning site with histidine tag, the internal sequences of malE and malG for homologous recombination, and the entire region of pGEM-T vector, except for the multiple cloning region, for propagation in E. coli. For stable expression of the target gene, an α-glucosidase-producing strain of S. acidocaldarius was generated employing this vector. The malA gene (saci_1160) encoding an α-glucosidase from S. acidocaldarius fused with the glutamate dehydrogenase (gdhAsaci) promoter and leader sequence was ligated to pINEX to generate pINEX_malA. Using the “pop-in” and “pop-out” method, the malA gene was inserted into the genome of MR31 and correct insertion was verified by colony PCR and sequencing. This strain was grown in YT medium without uracil and purified by His-tag affinity chromatography. The α-glucosidase activity was confirmed by the hydrolysis of pNPαG. The pINEX vector should be applicable in delineating gene functions in this organism.
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참고문헌 (21)
참고문헌 신청
Albers SV / 2008 / Conditions for gene disruption by homologous recombination of exogenous DNA into the Sulfolobus solfataricus genome / Archaea 2 : 145 ~ 149
Aucelli T / 2006 / A spreadable, non-integrative and high copy number shuttle vector for Sulfolobus solfataricus based on the genetic element pSSVx from Sulfolobus islandicus
Berkner S / 2007 / Small multicopy, non-integrative shuttle vectors based on the plasmid pRN1for Sulfolobus acidocaldarius and Sulfolobus solfataricus, model organisms of the
Berkner S / 2010 / Inducible and constitutive promoters for genetic systems in Sulfolobus acidocaldarius / Extremophiles 14 : 249 ~ 259
Bradford MM. / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 72 :
Aucelli T / 2006 / A spreadable, non-integrative and high copy number shuttle vector for Sulfolobus solfataricus based on the genetic element pSSVx from Sulfolobus islandicus
Berkner S / 2007 / Small multicopy, non-integrative shuttle vectors based on the plasmid pRN1for Sulfolobus acidocaldarius and Sulfolobus solfataricus, model organisms of the
Berkner S / 2010 / Inducible and constitutive promoters for genetic systems in Sulfolobus acidocaldarius / Extremophiles 14 : 249 ~ 259
Bradford MM. / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 72 :
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