| Product |
pUC18
&19 DNA |
| Description |
PUC18 &19 DNA are isolated from E.coli
(dam+, dcm+) and purified using alkalin method by phenol/chloroform
extraction. The molecule is a double- stranded circle,
2686 base pairs in length. |
| Aditional
information |
· CAP protein binding
site - 591-554 (compl. strand)
· mRNA (Lac
Z) starts at nt position 507 (compl. strand)
·
lac repressor binding site - 507-487 (compl. strand)
pUC18 and pUC19 vectors are small, high copy number, E
coli plasmids, 2686 bp in length. They are identical except
that they contain multiple cloning sites (MCS) arranged
in opposite orientation. pUC18/ 19 plasmids contain:
1- The pMB1 replicon rep responsible for the replication
of plasmid (source-plasmid pBR322). The high copy number
of pUC plasmid is a result of the lack of the rop gene
and a single point mutation in the replicon rep of pMB1.
2- The bla gene, coding for ß-lactamase, that confers
resistance to ampicillin (source-plasmid pBR322). It differs
from that of pBR322 by two point mutations.
3- The region of E. coli lac operon containing a CAP protein
binding site, promoter Plac, lac repressor binding site
and the 5'-terminal part of the lac Z gene encoding the
N-terminal fragment of ß-galactosidase (source-M13mp18/19).
This fragment, whose synthesis can be induced by IPTG,
is capable of intra-allelic æ-complementation with
a defective form of ß- galactosidase encoded by
the host (mutation Delta (lac Z)M15). In the presence
of IPTG, bacteria synthesize both fragments of the enzyme
and form blue colonies on media with X-Gal. Insertion
of DNA into the MCS located within the lacZ gene (codons
6-7 of lacZ are replaced by MCS) inactivates the N-terminal
fragment of ß-galactosidase and abolishes æ-complementation.
Bacteria carrying recombinant plasmids therefore give
rise to white colonies. |
| Storage |
-20°C |
| Storage
Buffer |
10mM Tris-HCl (pH 7.6)
and 1mM EDTA |
| Quality
Control |
More than 90% is in the
supercoiled form |
| Host |
E.coli HB101 |
|
