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Research Article

Isolation and Molecular Identification of Auxotrophic Mutants to Develop a Genetic Manipulation System for the Haloarchaeon Natrinema sp. J7-2

Table 1

Strains, plasmids, and amplification fragments used in this work.

Strains, plasmids, and amplification fragmentsDescriptionSource or reference

Strain
Natrinema sp. J7-2 Haloarchaeal strain isolated from a salt mineCCTCC AB91141, China [24, 25, 27]
Natrinema sp. J7-2-1Auxotrophic strain from J7-2, LeuThis work
Natrinema sp. J7-2-22Auxotrophic strain from J7-2, ArgThis work
Natrinema sp. J7-2-26Auxotrophic strain from J7-2, LysThis work
Natrinema sp. J7-2-52Auxotrophic strain from J7-2, ArgThis work
Escherichia coli DH5α  endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG 80dlacZM15 (lacZYA-argF)U169,  hsdR17(), [42]
Escherichia coli JM110rpsL thr leu thi lacY galK galT ara tonA tsx dam dcm glnV44 (lac-proAB) e14-[ traD36 lacI lacZM15] hsdR17() [43]
Plasmid
 pUC19Cloning vectorLab stock
 pMD19T-simpleSequence vectorPurchased from TaKaRa
 pUC19-KM1752pUC19 containing leuB gene with KpnI and MfeI from J7-2This work
 pUC19-HM1752pUC19 containing the ORF of leuB gene with HindIII and MfeI from J7-2This work
 pUC19-HM4049pUC19 containing dapD gene with HindIII and MfeI from J7-2This work
 pUC19-HM1752-amyH pUC19-HM1752 containing amyH gene from Haloarcula hispanica DSM 4426This work
 pUC19-HM1752-bgaH pUC19-HM1752 containing bgaH gene from Haloferax  alicantei This work
Amplification fragments
 KM1752leuB gene containing promoter from J7-2 with KpnI and MfeIThis work
 HM1752Amplification fragments of leuB gene from J7-2 with HindIII and MfeIThis work
 HM4049Amplification fragments of dapD gene from J7-2 with HindIII and MfeIThis work

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