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

Table 4

Molecular characterization of the auxotrophic mutants by analyzing relevant genes in the corresponding amino acid biosynthetic pathways.

Auxotrophic strains	Phenotype	Gene	Gene number	Notation	Length (bp)	Mutation status
J7-2-1	Leu^−	leuA	NJ7_G1272	2-Isopropylmalate synthase	831	None
		leuC	NJ7_G1748	3-Isopropylmalate dehydratase, large subunit	1388	None
		leuD	NJ7_G1749	3-Isopropylmalate dehydratase, small subunit	837	None
		leuB	NJ7_G1752	3-Isopropylmalate dehydrogenase	768	12 bp fragment deletion from 690 to 701 bp
J7-2-26	Lys^−	dapA	NJ7_G4051	Dihydrodipicolinate synthase	951	None
		dapF	NJ7_G4046	Diaminopimelate epimerase	1639	None
		lysA	NJ7_G4047	Diaminopimelate decarboxylase	1422	None
		dapD	NJ7_G4049	2,3,4,5-Tetrahydropyridine-2,6-dicarboxylate N-succinyltransferase	635	18 bp deletion from 583 to 600 bp
		dapB	NJ7_G4050	Dihydrodipicolinate reductase	987	None
Auxotrophic strains	Phenotype	Gene	Gene number	Notation	Length (bp)	Mutation Status
						J7-2-22	J7-2-52
J7-2-22; J7-2-52	Arg^−	argF	NJ7_G0258	Ornithine carbamoyltransferase	921	None	None
		argE	NJ7_G0259	N-Acetyl-ornithine/N-acetyl-lysine deacetylase	1110	None	None
		argD	NJ7_G0260	Acetylornithine aminotransferase	1155	None	None
		argB	NJ7_G0261	Acetylglutamate kinase	870	None	None
		argC	NJ7_G0262	N-Acetyl-gamma-glutamyl-phosphate reductase	1077	None	1051 bp: G-A(GGG-GAG)
		argX	NJ7_G0263	Ribosomal protein S6 modification protein	1043	None	None
		argW	NJ7_G0264	Putative biosynthetic carrier protein
		argH	NJ7_G0265	Argininosuccinate lyase	1539	None	None
		argG	NJ7_G0266	Argininosuccinate synthase	1242	None	None