pMN20植物激活标签载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pMN20

pMN20植物激活标签载体

BioVector NTCC质粒载体菌种细胞基因保藏中心                      

Used to confirm activation-tagged genes. Contains multiple cloning sites adjacent to the same tetramerized CaMV 35S enhancers as in the activation-tagging vectors from this article.

Kanamycin gene for plant selection on MS media.

Plasmid selection in E. coli and in A. tumefaciens is spectinomycin (100 mg/l).

The four enhancer repeats in the construct may be unstable in E. coli and Agrobacterium if stored at +4'C for extended time. Check by PCR with T7 (or M13-20) oligo and this one derived from the RB sequence: 5' acc cgc caa tat atc ctg 3'. This should give 1.4 kb band. Note that these oligos will not work in a transgenic plant, as the RB sequence is not transfered. Using this test, we found that after 1-2 weeks in a fridge the Agrobacterium strain loses on average one copy of a repeat, and after a month in a fridge there is only one copy left. It is a good idea to use a freshly streaked colony from a good -70'C glycerol stock for every infiltration.
Vector backbone pPZP212
(Search Vector Database)
Backbone manufacturer Hajdukiewicz et al 1994
Backbone size w/o insert (bp) 11073
Vector type Plant Expression
Selectable markers kanamycin
GROWTH IN BACTERIA

Bacterial Resistance(s) Spectinomycin
Growth Temperature 37°C
Growth Strain(s) DH5alpha
Growth instructions Plasmid grows in standard E. coli at 37 degrees, but the four enhancer repeats may be unstable in E. coli and A. tumefaciens if stored at 4'C for extended time. See comments.
Copy number High Copy
GENE/INSERT

Gene/Insert name CaMV 35S enhancers
CLONING INFORMATION

Cloning method Restriction Enzyme
5′ cloning site See paper and map. (not destroyed)
3′ cloning site See paper and map. (not destroyed)
5′ sequencing primer M13 reverse

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
电话：+86-010-53513060
网址：www.biovector.net      [Supplier来源]  http://www.biovector.net