Helicobacter pylori HopE and HopV porins present scarce expression among clinical isolates

Table 1 Primer sequences used for amplification and sequencing of H. pylori hopE and hopV genes

Name	5'→3' sequence	Restriction sites1
HopV12	GGGCCCATATGCTCAATTTTATGACAAAGAAGAAAAATAGAATGC	NdeI
HopV22	GGATCCCATGGTTAAAAATCCCTCAAGTAACTGATTTG	BamHI
HopE12	GGCGCCATGGAATTTATGAAAAAGTTTGTAGCTTTAGG	NcoI
HopE22	CGCGAAGCTTTTAAAAAGTGTAGTTATACCCTAAATAAAG	HindIII
HopE113	GCAAGTGGTTTGGTTTTAGAG	-
HopE223	ACCATATCCAACTGGATTTT	-
HopV113	GGCGTGGGGTTAGATACGCTG	-
HopV223	ACCATGTTTTCTTTATTCAC	-
HopVint3	ATGCGTTATTATGGGTTTTTTGACT	-
pETT7d4	TAATACGACTCACTATAGGG	-
pETT7r4	GCTAGTTATTGCTCAGCGG	-

¹The restriction sites included in primer sequences and used for ligation to plasmid vectors are shown underlined;

²External primers used to clone porin genes;

³Internal primers used for gene sequencing and also for confirmatory PCR reactions for those cases in which primers derived from 5’ and 3’ gene ends failed to raise PCR products;

⁴Vector primers for 5’ and 3’ end gene sequencing.

Table 2 Number of strains presenting genotypes and corresponding phenotypes indicating the presence of hopE (E) and hopV (V) genes and their expression in 130 H. pylori strains isolated from infected patients from 6 Chilean cities

Genotypes1 (E/V)	No. of strains	Phenotypes2 (E/V)	No. of strains	No. of strains with different E/V phenotypes per city3
				IQ	VA	SA	LA	VL	PA
(+/+)	52	(+/+)	5	4	-	1	-	-	-
		(+/-)	12	4	-	5	1	-	2
		(-/+)	-	-	-	-	-	-	-
		(-/-)	35	15	3	3	1	7	6
(+/- )	9	(+/-)	-	-	-	-	-	-	-
		(-/-)	9	2	-	-	-	5	2
(-/+)	30	(-/+)	4	1	-	2	-	-	1
		(-/-)	26	11	-	4	-	2	9
(-/-)	39	(-/-)	39	11	2	11	2	7	6
Totals	130	Totals	130	48	5	26	4	21	26

¹Presence of hopE (E) and/or hopV (V) genes in Chilean H. pylori isolates determined by PCR amplifications as described in Methods using purified DNA templates from single colonies collected from 69 patients of the indicated cities. (+): gene presence; (-): no detection;

²HopE and or HopV expression assayed by Western blotting (see Methods). (+): detection; (-): no detection;

³Total number of strains with the assigned phenotype per city. IQ: Iquique; VA: Valparaiso; SA: Santiago; LA: Los Angeles; VL: Valdivia; PA: Punta Arenas. The number of patients per city was IQ = 21, VA = 4, SA = 12, LA = 3, VL = 5, PA = 15. Number of strains isolated per patient ranged between 1 and 5.