True. These bacteria contain a large plasmid named Ti plasmid, which is causative agent of crown gall disease in plants. A shuttle vector as in claim 6, having a structural gene foreign to the pTAR plasmid inserted sufficiently close to the T-DNA so that said structural gene can be transferred to a Ti plasmid by homologous recombination. Agrobacterium -mediated plant transformation has been used widely, but there are plants that are recalcitrant to this type of transformation. Title: Ti plasmid derived vector system 1 Lecture 3. false. pUCD400. Modified shuttle Ti plasmids were extracted from A, tumefaciens strains by the modified alkaline sodium dodecyl sulfate method and purified by ethidium bromide-CsCl gradient ultracentrifugation.

Binary vector is otherwise called as Shuttle vector. The gene of interest is inserted into the vector plasmid by means of a restriction/ligation event carried out by the use of suitable restriction enzymes. an E.coli - A. tumifaciens shuttle vector o r a . DEVELOPMENT OF BINARY VECTOR SYSTEMS. A shuttle vector as in claim 3, further including at least a portion of the T-DNA region derived from a Ti plasmid. The recombinant BPV is constructed by ligating our gene of interest and BPV vector (69%) onto the pBR 322 plasmid, thus generating the shuttle vector containing plasmid ori site and virus replication sequences. These shuttle vectors are multiplied in E. coli cells first and then they are … The Ti plasmid isolated from Agrobacterium can be used to insert DNA into any type of plant. This transformation method uses bacterial strains harboring a modified tumor-inducing (Ti) plasmid that lacks the transfer DNA (T-DNA) region (disarmed Ti plasmid). Shuttle Vectors ... the Ti plasmid is used in plant pathology to develop resistance in plants against diseases such as holcus spot on leaves and crown gall tumors. T DNA is inserted into the pBR 322 vector which results in formation of a shuttle vector. A shuttle vector is a plasmid that is used to move pieces of DNA among organisms, such as bacterial, fungal, and plant cells. In transferring gene of interest to Ti plasmid, an intermediate vector is used such as pBR 322. It is desirable to develop strains that can broaden the host range.

Essentially binary vector contains both border sequences, marker genes and many unique restriction sites for the insertion of target gene between T-DNA borders. It is possible to insert a desired DNA sequence (gene) into the T-DNA region (of Ti-plasmid),and then use A. tumefaciens to deliver this gene into the genome of plant cell. This shuttle vector can replicate in E. coli and in Agrobacterium. Binary vector can be maintained in Agrobacterium without the process of co-integration into the Ti plasmid.