Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 May 14;10:923. doi: 10.3389/fimmu.2019.00923

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2019 Kilpeläinen, Saubi, Guitart, Moyo, Wee, Ravi, Hanke and Joseph.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

Construction of the BCG.HIVconsv1&2^2auxo.int vaccines. (A) Schematic of the construction of the tHIVconsvX immunogens (28). (B) The HIVconsv1 and HIVconsv2 (gray) synthetic sequences, were BCG codon optimized and fused to the 19-kDa lipoprotein signal sequence and inserted into the integrative p2auxo.HIVconsv1^int and p2auxo.HIVconsv2^int E. coli-mycobacterial shuttle plasmids, respectively. These shuttle vectors contain P α-Ag (in red), Mycobacterium tuberculosis α-antigen promoter, P HSP60, heat shock protein 60 gene promoter. The glyA and LysA (in yellow) complementing genes function as an antibiotic-free selection system in the auxotrophic strains of E. coli M15ΔglyA and BCG ΔLys, respectively. At the bottom, the process of plasmid integration in the BCG genome, based on the integration system of the L5 phage, is shown. The recognition occurs between the attachment sites AttP and AttB of the plasmid and mycobacterial genome, respectively (Att: attachment sites, in violet). (C) PCR analysis of attR and attL DNA regions of BCG.HIVconsv1^2auxo.int (left) and BCG.HIVconsv2^2auxo.int (right) colonies. Lanes 2 and 10: BCG wild type; Lanes 3 and 11: BCG.HIVconsvX^2auxo.int clone1; Lanes 4 and 12: BCG.HIVconsvX^2auxo.int clone 2; Lanes 5 and 13: BCG.HIVconsvX^2auxo.int clone 3; Lanes 6 and 14: BCG.HIVconsvX^2auxo.int clone 4; Lanes 7 and 15: negative control, distilled water; and lanes 1,8,9 and 16: molecular weight marker. (D) Western blot of BCG.HIVconsv1^2auxo.int and BCG.HIVconsv2^2auxo.int cell lysates. Lane 1: BCG wild type (negative control); Lane 2: BCG.HIVconsv1^2auxo.int working vaccine stock; Lane 3: BCG.HIVconsv2^2auxo.int working vaccine stock; Lane 4: BCG.Ø^2auxo.int; Lane 5: Molecular weight marker.