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. Author manuscript; available in PMC: 2015 Jun 16.
Published in final edited form as: Virology. 2009 Mar 20;387(2):388–394. doi: 10.1016/j.virol.2009.02.024

Figure 4. Construction of the V2 chimeric Envs.

Figure 4

(A) Env sequences encoding donor V2 domains (MBA and LEA) were amplified by PCR using specific primers that anneal at both ends of the V2 domain. (B) Env sequences encoding recipient env backbones within the pCR2.1 vector (pCR2.1–envΔV2MBA and pCR2.1–envΔV2LEA) were amplified by PCR using specific primers that anneal to regions adjacent to the V2 primers. A 5’ phosphate group was added to these primers for ligation to the V2 amplicon. (C) Sequences encoding the V2 domains and recipient backbones were blunt-end ligated to generate chimeric env genes in pCR2.1 (pCR2.1–V2MBA-envΔV2LEA and pCR2.1–V2LEA-envΔV2MBA).