pcombia载体图谱.doc

pCAMBIA1305.1 and pCAMBIA1305.2 This vector series is based on a new reporter gene GUSPlus(. The novel reporter enzyme has improved enzymatic characteristics, and can be more efficiently secreted than E. coli GUS when fused with a signal peptide.  pCAMBIA1305.1 and pCAMBIA1305.2 CAMBIA’s GUSPlus( is a new reporter gene isolated from Staphylococcus sp with superior properties to E.coli GUS Better catalytic activity for more rapid detection of GUS activity. Greater stability at 60(C and in the presence of fixatives such as formaldehyde and glutaraldehyde for robust, cell-specific detection of gene activity in situ. A version with the rice glycine-rich protein signal peptide for extracellular secretion providing rapid, in vivo gus assays. GUSPlus( has been tested extensively in transgenic rice, and the gene has been codon optimised for high expression in plants. The version in pCAMBIA1305.2 with a signal peptide provides an alternative to GFP, offering sensitive in vivo observation of gene activity without the need for UV detection. GUSPlus( is available in two vector formats: pCAMBIA1305.1 (Genbank acc nr AF354045). This vector is based on pCAMBIA1300 in which E. coli gusA has been replaced by GUSPlus(. It is compact binary vector with a pBR322 ori for high copy replication in E.coli and a broad host range ori for low copy, stable replication in Agrobacterium. It contains the hygromycin resistance gene (hpt II) for selection in plants. The GUSPlus( gene contains the intron from the castor bean catalase gene to ensure detection of plant-specific glucuronidase expression. pCAMBIA1305.2 (Genbank acc nr AF354046). Same as pCAMBIA1305.1 except for the presence of the rice glycine-rich protein signal peptide sequence (GRP) inserted between the Nco I and Bgl II sites. General features pCAMBIA 1305.1 pCAMBIA 1305.2 pUC18 multiple cloning site Blue/white (-galactosidase screening High copy plasmid in E.coli Triparental mating feasibility GUSPlus( replacement of gusA