Abstract
The gateway technology developed by Invitrogen is very useful for construction of the expression vectors for a target gene. However, the present destination vectors developed by VIB/Gent for expressing target genes in plants only contain 35S promoter. The expressed proteins driven by 35S are localized in cytoplasm. Thus, the present gateway technology can not be used to construct plant expression vectors for localization of target proteins into chloroplasts. We converted Xmn I site in gateway entry vector, pENTR-2B, as HindIII site and then inserted Rubisco small subunit promoter (PrbcS) from tomato and GFP gene into pENTR-2B for generation of a gateway entry vector (pENTR-PrbcS-T-GFP). Here, we showed that pENTR-PrbcS-T-GFP can be applied to generate the entry vector for any target gene by replacement of GFP with a target gene. The plant expression vectors for localization of target proteins into chloroplasts can be made quickly by gateway technology.
