vector containing primer sites useful for sequencing
vector for studying DNA-protein interactions
Comments
One of three vectors (ATCC 87121-87123) designed for studying the amount of bending induced at a DNA binding site due to DNA-protein interactions. The three vectors differ only in the available cloning sites.
Vector constructed by insertion of the pBend2 polylinker into pBluescriptSK- at the EcoRI-HindIII sites followed by destruction of the Bluescript XbaI and SalI sites.
Target DNA binding sites can be cloned into the vector and an array of fragments can be generated (by restriction digestion with different enzymes) that are equal in length, but differ in the position of the DNA binding site along the fragment.
The reduced mobility of different protein-fragment complexes can then by analyzed by gel electrophoresis to determine the degree of bending induced by protein binding.
Restriction digests of the clone give the following sizes (kb): BglI--1.6, 1.3, 0.32, 0.24; EcoRI/HindIII--3.0, 0.29; XbaI--3.2.
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References
Kim J, et al. Bending of DNA by gene-regulatory proteins: construction and use of a DNA bending vector. Gene 85: 15-23, 1989. PubMed: 2533576
Zwieb C, Adhya SImproved plasmid vectors for the analysis of protein-induced DNA bendingIn: Zwieb C, Adhya SMethods in Molecular Biology, Vol 30: DNA-Protein interactions: Principles and ProtocolsTotowa, NJThe Humana Press, Inc.pp. 281-294, 1994
