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 2–24	2: Asp	3: Cys	4: Ile
5: Gly	6: Glu	7: Val	8: Ser
9: Ala	10: His	11: Tyr	12: Phe
13: Met	14: Thr	15: Met	16: Trp
17: Pro	18: Leu	19: Lys	20: cystine
21: Arg	22: Asp	23: Asn	24: Gln
Group 25: containing vitamins from 26 to 35	26: vitamin H	27: vitamin C	28: vitamin B6
29: vitamin B3	30: vitamin B5	31: vitamin B4	32: vitamin B2
33: vitamin B9	34: vitamin B12	35: vitamin B1
Group 36: containing dNTPs from 37 to 43	37: dATP	38: dGTP	39: dTTp
40: dCTP	41: dUTP	42: xanthine	43: hypoxanthine

(b) Different combination groups containing amino acids

	Group A	Group B	Group C	Group D	Group E
Group F	9: Ala	21: Arg	23: Asn	22: Asp	3: Cys
Group G	6: Glu	24: Gln	5: Gly	10: His	4: Ile
Group H	18: Leu	19: Lys	15: Met	12: Phe	17: Pro
Group I	8: Ser	14: Thr	16: Trp	11: Tyr	7: Val

(c) Different combination groups containing vitamins

	Group I	Group II	Group III
Group IV	32: vitamin B2	33: vitamin B9	31: vitamin B4	35: vitamin B1
Group V	28: vitamin B6	27: vitamin C	29: vitamin B3	35: vitamin B1
Group VI	30: vitamin B5	34: vitamin B12	26: vitamin H	35: vitamin B1

(d) Different combination groups containing purine and pyrimidine

	Group X1	Group X2	Group X3
Group X4	41: dUTP	37: dATP	39: dTTP
Group X5	42: xanthine	43: hypoxanthine	40: dCTP
	38: dGTP	38: dGTP	38: dGTP

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

Supplied nutrients in MM medium	J7-2-1	J7-2-22	J7-2-26	J7-2-52
MM medium	N^a	N	N	N
MGM medium	Y^b	Y	Y	Y
Group ALL	Y	Y	Y	Y
Group 1	Y	Y	Y	Y
Group 25	N	N	N	N
Group 36	N	N	N	N
Group A	Y	N	N	N
Group B	N	Y	Y	Y
Group F	N	Y	N	Y
Group H	Y	N	Y	N
Group I	N	I^c	N	N
18	Y	N	N	N
19	N	N	Y	N
21	N	Y	N	Y
Other nutrients	N	N	N	N

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