pBP96 (ATCC® 77084)

Applications: YI-type (integrating) shuttle vector  /  Depositors: RH Reeves

Permits and Restrictions

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Designations pBP96
Depositors RH Reeves
Biosafety Level 1
Vector Information
Size (kb): 4.5999999046325680
Vector: pBP96 (plasmid)
Construction: BLUR8, HIS3
Marker(s):ampR,HIS3
Construct size (kb): 4.599999904632568
Features: marker(s): ampR, HIS3
replicon: pMB1
Applications
YI-type (integrating) shuttle vector
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--3.0, 1.6; EcoRI--4.6; PstI--3.3, 1.3.
Digestion with BamHI removes the plasmid sequences from the Alu ends for targeting integration.
Transformation into pools from a YAC library derived from a human-mouse somatic cell hybrid, and selection for His+, resulted in an approximate 50-fold enrichment for Alu-containing YACs.
Larger YACs were preferentially selected. Some YACs demonstrated loss of Alu-containing restriction fragments suggesting internal deletions resulting when the vector targeted two Alu sequences in the YAC.
YI-type shuttle integrative vector targeting human Alu sequences in YAC libraries derived from somatic cell hybrids.
Constructed by inserting a blunt-ended 1.2 kb fragment containing S. cerevisiae HIS3 into the blunted MscI within the Alu sequence of BLUR8.
The order of the major features in this plasmid is: bla - BamHI - Alu - HIS3 - Alu - BamHI.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Pavan WJ, Reeves RH. Integrative selection of human chromosome-specific yeast artificial chromosomes. Proc. Natl. Acad. Sci. USA 88: 7788-7791, 1991. PubMed: 1881915

Reeves RH, et al. Yeast artificial chromosome modification and manipulation. Methods Enzymol. 216: 584-603, 1992. PubMed: 1336105

Roger H Reeves, personal communication

Shipped freeze-dried
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