P gem3 z example of puc vector

P gem3 z example of puc vector

1. pGEM3-z By:- Vivek kuma5r M.Sc Microbiology Bangalore University

2. PLASMIDS A Plasmid is a self replicating,double stranded , extrachromosomal DNA’s maintained as independent molecules by the most of the bacterial genera. LEDERNBERG IN 1952 C oined the term “PLASMID”.

3. CLONING VECTORS BASED ON E.Coli PLASMIDS ex: pUC 8 plasmid pUC8 plasmid is derived from pBR322, remaining only the replication origin and the ampR gene. All coding sites are cloning sites are clustered into a short segment of the Lac Z gene carried by pUC 8.

4. Used in invitro transcription and protein expression. pGEM3-Z is having similarity with a pUC vector because of the presence of the ampR genes and Lac Z gene containing a cluster of restriction sites.

5. advantages High copy number. Identification can be achieved by single step by plating on agar medium containing amplicillin and X-gal . Time consumption is less. pUC vectors carry different combinations of restrictions site and show greater flexibility of DNA fragment that can be cloned. Clustering of restriction sites that show greater flexibility allows a DNA fragment with two different sticky ends to be cloned without involving linker attachment. DNA cloned into a member of the pUC series can be transferred directly to its M13 mp counterpart, because of the same restriction sites clusters and it can be analyzed by DNA sequencing or in vitro Mutagenesis.

6. THANK YOU