Generation of comprehensive transposon insertion mutant library for the model archaeon, Haloferax volcanii, and its use for gene discovery

Table 5 Amino acid auxotrophic mutants

Mutant	Auxotrophy	Integration site; orientation ^a	Gene	Protein
H295/6-78	Arginine	43,942; +	HVO_0044, argB	Acetylglutamate kinase
H295/6-100	Arginine	44,698: +	HVO_0045, argC	N-acetyl-gamma-glutamyl-phosphate reductase
H295/12-27	Arginine	44,905; -	HVO_0045, argC	N-acetyl-gamma-glutamyl-phosphate reductase
H295/10-36	Arginine	43,144; +	HVO_0043, argD	Acetylornithine aminotransferase
H295/17-48	Arginine	2,374,426; -	HVO_2508, carA	Carbamoyl-phosphate synthase small subunit
H295/13-54	Histidine	1,198,248; +	HVO_0431	HAD-superfamily hydrolase
H295/12-18	Histidine	1,198,248; +	HVO_0431	HAD-superfamily hydrolase
H295/21-77	Proline	786,914; +	HVO_0869	Glutamate synthase [NADPH] large chain
H295/24-65	Proline	1,051,586; -	HVO_1153	Hypothetical protein
H295/12-1	Tyrosine	1,198,243; +	HVO_1312	Prephenate dehydrogenase
H295/16-1	Isoleucine	578,651; -	HVO_0644, leuA1	2-isopropylmalate synthase/(R)-citramalate synthase
H295/24-63	Phenylalanine	399,114; -	HVO_0449, pheA1	Prephenate dehydratase
H295/9-8	Valine, Leucine, Isoleucine	1,374,516; -	HVO_1506, ilvC	Ketol-acid reductoisomerase
H295/4-26	Valine, Leucine, Isoleucine	1,375,827; +	HVO_1508, ilvB1	Acetolactate synthase large subunit
H295/12-44	Glycine, Serine, Threonine	2,805,618; +	HVO_2969, thrC3	Threonine synthase

^aThe first change compared to H. volcanii chromosomal sequence (NC_013967.1) is indicated as the transposon integration site. The orientation is denoted as trpA direction compared to H. volcanii chromosome (5′ to 3′): +, same direction; -, opposite direction.

ISSN: 1741-7007