Leishmania major large RAB GTPase is highly immunogenic in individuals immune to cutaneous and visceral leishmaniasis

Table 2 Primers used for cloning of LmlRAB and LmlRABC and plasmid constructions

Name	Type	Primer sequences	Primer sequences	Restriction site
F-Ras1a	Forward	rlmlrab	5′-GCCCATATGAGCTCAACTGGTCAGCATG-3′	NdeI
F-Ras3a	Forward	rlmlrabC	5′GCCCATATGATCATCGCCGATGTGTCG-3′	NdeI
R-Ras3b	Reverse	rlmlrab/rlmlrabC	5′-GCCGCGGCCGCCCAGTACGCAGCAGTTGC-3′	NotI

Reverse and forward primers containing at their 5′ end a restriction site (underlined) were used to amplify the full length LmlRAB ORF and its carboxyl part LmlRABC. The amplified double-stranded DNA were first digested with NdeI and NotI and then inserted into the corresponding cloning sites in the pET-22b + vector (Novagen)

ISSN: 1756-3305