Construction of Expression Shuttle Vectors for the Haloarchaeon Natrinema sp. J7 Based on Its Chromosomal Origins of Replication

Table 3

Characteristics of predicted oriCs and replication regions used for shuttle vector construction.

(a) Characteristic of predicted oriCs

OriCs	Locations	Number of ORB elements	GC content (%)	Adjacent to cdc6 gene
1	120445–121114	2	51.27%	Y
2	122480–123045	1	60.30%	Y
3	399506–400667	3	56.44%	N
4	434294–435598	8	54.87%	Y
5	1277983–1279633	5	59.37%	Y
6	1280840–1281049	1	66.67%	Y
7	2211660–2211903	2	60.51%	Y
8	2213131–2213664	2	62.89%	Y
9	2573530–2574443	1	55.78%	Y
10	2860326–2861584	7	52.74%	N
11	3151118–3151637	3	60.17%	Y
12	3153862–3154084	1	68.61%	Y

(b) Characteristic and replication ability of the shuttle vectors

Shuttle vectors	ARSs	OriCs contained	Chromosomal locations of ARSs (positions)	Containing; cdc6 gene	Transformation efficiency
pFJ1	1	1, 2	120,345–123,145	Y	() × 10^3
pFJ2	2	4	434,194–437,483	Y	0
pFJ3	3	5, 6	1,277,883–1,281,049	Y	(3.9 ± 0.4) × 10^1
pFJ4	4	7, 8	2,211,560–2,213,764	Y	(2.1 ± 0.1) × 10^3
pFJ5	5	9	2,572,200–2,574,543	Y	<10
pFJ6	6	10	2,860,000–2,862,000	N	(2.2 ± 0.5) × 10^3
pFJ7	7	3	399,506–400,667	N	<10

-forming units/μg DNA.