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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 3

Different combination groups of growth factors used in this work to determine the auxotrophic phenotype of the four mutation strains.
(a) Single growth factors used in this work and groups containing all of the growth factors belonged to the same type

Group 1: containing amino acids from 2–242: Asp3: Cys4: Ile
5: Gly6: Glu7: Val8: Ser
9: Ala10: His11: Tyr12: Phe
13: Met14: Thr15: Met16: Trp
17: Pro18: Leu19: Lys20: cystine
21: Arg22: Asp23: Asn24: Gln

Group 25: containing vitamins from 26 to 3526: vitamin H27: vitamin C28: vitamin B6
29: vitamin B330: vitamin B531: vitamin B432: vitamin B2
33: vitamin B934: vitamin B1235: vitamin B1

Group 36: containing dNTPs from 37 to 4337: dATP38: dGTP39: dTTp
40: dCTP41: dUTP42: xanthine43: hypoxanthine

(b) Different combination groups containing amino acids

Group AGroup BGroup CGroup DGroup E

Group F9: Ala21: Arg23: Asn22: Asp3: Cys
Group G6: Glu24: Gln5: Gly10: His4: Ile
Group H18: Leu19: Lys15: Met12: Phe17: Pro
Group I8: Ser14: Thr16: Trp11: Tyr7: Val

(c) Different combination groups containing vitamins

Group IGroup IIGroup III

Group IV32: vitamin B233: vitamin B931: vitamin B435: vitamin B1
Group V28: vitamin B627: vitamin C29: vitamin B335: vitamin B1
Group VI30: vitamin B534: vitamin B1226: vitamin H35: vitamin B1

(d) Different combination groups containing purine and pyrimidine

Group X1Group X2Group X3

Group X441: dUTP37: dATP39: dTTP
Group X542: xanthine43: hypoxanthine40: dCTP
38: dGTP38: dGTP38: dGTP

(e) The growth of the four auxotrophic strains on MM medium supplied with different nutrients

Supplied nutrients in MM mediumJ7-2-1J7-2-22J7-2-26J7-2-52

MM mediumNaNNN
MGM mediumYbYYY
Group ALLYYYY
Group 1YYYY
Group 25NNNN
Group 36NNNN
Group AYNNN
Group BNYYY
Group FNYNY
Group HYNYN
Group INIcNN
18YNNN
19NNYN
21NYNY
Other nutrientsNNNN

N: the growth of the auxotrophs was not detected. bY: the growth of the auxotrophs was detected. cI: an inhibition zone was detected.

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