Stable nuclear transformation of Chlamydomonas reinhardtii with a Streptomyces rimosus gene as the selective marker

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Stable nuclear transformation of Chlamydomonas reinhardtii with a Streptomyces rimosus gene as the selective marker. / Sizova, Irina A.; Lapina, Tatyana V.; Frolova, Olga N.; Alexandrova, Nelly N.; Akopiants, Konstantin E.; Danilenko, Valery N.

в: Gene, Том 181, № 1-2, 28.11.1996, стр. 13-18.

Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование

Author

Sizova, Irina A. ; Lapina, Tatyana V. ; Frolova, Olga N. ; Alexandrova, Nelly N. ; Akopiants, Konstantin E. ; Danilenko, Valery N. / Stable nuclear transformation of Chlamydomonas reinhardtii with a Streptomyces rimosus gene as the selective marker. в: Gene. 1996 ; Том 181, № 1-2. стр. 13-18.

BibTeX

@article{5228dc7643c94db4861cf73d25876ae7,

title = "Stable nuclear transformation of Chlamydomonas reinhardtii with a Streptomyces rimosus gene as the selective marker",

abstract = " The aminoglycoside 3'-phosphotransferase typeVIII (APHVIII) encoding gene (aphVIII) from Streptomyces rimosus was introduced by glass-bead high-efficiency transformation into the nuclear genome of green unicellular alga Chlamydomonas reinhardtii for induction of transformants resistant to aminoglycoside antibiotics. The aphVIII structural sequence was flanked by S. rimosus regulatory sequences which failed to direct expression in C. reinhardtii. The pSU937 plasmid containing these sequences was able to transform C. reinhardtii strain cw15 arg7-8 mt + for paromomycin resistance (Pm(R)) at a frequency (1.3-1.9) x 10 -7 , probably as a result of in vivo gene fusion and expression of the aphVIII gene from regulatory elements of nuclear DNA. Evidence for the real C. reinhardtii transformation includes blot-hybridization with a probe specific for aphVIII and demonstration of APHVIII enzyme activity in crude cell extracts of transformants. Integrated Streptomyces DNA sequences, APHVIII enzyme activity and the aminoglycoside-resistance phenotype were stable through mitosis in the presence and absence of selection. The Pm(R) phenotype was inherited by the meiotic progeny of transformants from crosses with wild-type strains. ",

keywords = "Aminoglycoside 3'-phosphotransferase, Expression, Foreign gene, Gene fusion",

author = "Sizova, {Irina A.} and Lapina, {Tatyana V.} and Frolova, {Olga N.} and Alexandrova, {Nelly N.} and Akopiants, {Konstantin E.} and Danilenko, {Valery N.}",

year = "1996",

month = nov,

day = "28",

doi = "10.1016/S0378-1119(96)00384-8",

language = "English",

volume = "181",

pages = "13--18",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

number = "1-2",

}

RIS

TY - JOUR

T1 - Stable nuclear transformation of Chlamydomonas reinhardtii with a Streptomyces rimosus gene as the selective marker

AU - Sizova, Irina A.

AU - Lapina, Tatyana V.

AU - Frolova, Olga N.

AU - Alexandrova, Nelly N.

AU - Akopiants, Konstantin E.

AU - Danilenko, Valery N.

PY - 1996/11/28

Y1 - 1996/11/28

N2 - The aminoglycoside 3'-phosphotransferase typeVIII (APHVIII) encoding gene (aphVIII) from Streptomyces rimosus was introduced by glass-bead high-efficiency transformation into the nuclear genome of green unicellular alga Chlamydomonas reinhardtii for induction of transformants resistant to aminoglycoside antibiotics. The aphVIII structural sequence was flanked by S. rimosus regulatory sequences which failed to direct expression in C. reinhardtii. The pSU937 plasmid containing these sequences was able to transform C. reinhardtii strain cw15 arg7-8 mt + for paromomycin resistance (Pm(R)) at a frequency (1.3-1.9) x 10 -7 , probably as a result of in vivo gene fusion and expression of the aphVIII gene from regulatory elements of nuclear DNA. Evidence for the real C. reinhardtii transformation includes blot-hybridization with a probe specific for aphVIII and demonstration of APHVIII enzyme activity in crude cell extracts of transformants. Integrated Streptomyces DNA sequences, APHVIII enzyme activity and the aminoglycoside-resistance phenotype were stable through mitosis in the presence and absence of selection. The Pm(R) phenotype was inherited by the meiotic progeny of transformants from crosses with wild-type strains.

AB - The aminoglycoside 3'-phosphotransferase typeVIII (APHVIII) encoding gene (aphVIII) from Streptomyces rimosus was introduced by glass-bead high-efficiency transformation into the nuclear genome of green unicellular alga Chlamydomonas reinhardtii for induction of transformants resistant to aminoglycoside antibiotics. The aphVIII structural sequence was flanked by S. rimosus regulatory sequences which failed to direct expression in C. reinhardtii. The pSU937 plasmid containing these sequences was able to transform C. reinhardtii strain cw15 arg7-8 mt + for paromomycin resistance (Pm(R)) at a frequency (1.3-1.9) x 10 -7 , probably as a result of in vivo gene fusion and expression of the aphVIII gene from regulatory elements of nuclear DNA. Evidence for the real C. reinhardtii transformation includes blot-hybridization with a probe specific for aphVIII and demonstration of APHVIII enzyme activity in crude cell extracts of transformants. Integrated Streptomyces DNA sequences, APHVIII enzyme activity and the aminoglycoside-resistance phenotype were stable through mitosis in the presence and absence of selection. The Pm(R) phenotype was inherited by the meiotic progeny of transformants from crosses with wild-type strains.

KW - Aminoglycoside 3'-phosphotransferase

KW - Expression

KW - Foreign gene

KW - Gene fusion

UR - http://www.scopus.com/inward/record.url?scp=0030605237&partnerID=8YFLogxK

U2 - 10.1016/S0378-1119(96)00384-8

DO - 10.1016/S0378-1119(96)00384-8

M3 - Article

C2 - 8973302

AN - SCOPUS:0030605237

VL - 181

SP - 13

EP - 18

JO - Gene

JF - Gene

SN - 0378-1119

IS - 1-2

ER -

ID: 45860934