Question

Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed the following set of restriction digests on a newly isolated plasmid, pBLA230. The reaction they carries out, along with the fragment obtained in single and double digest reactions were: Enzyme Eragment Lengths obtained: Hpal HindITI 26 kb 13 kb, 6 kb, 4 kb, 3 kb Hpal + HindITI 7 kb, 6 kb (2), 4 kb, 3 kb pBLA230 (26kb) Using this information, construct a restriction map of PBLA230.

Answer 1

1) there are 3 BamHI sites, two BamHI sites are separated by 6 kb and the other BamHI site is placed 2 kb from one BamHI site. so three bands are obtained by the digestion of BamHI.

2kb, 6kb, and 12kb.

there is only one EcoRI site, so EcoRI will cut and linearize the plasmid, so the size of the band obtained with the EcoRI digestion is 20kb.

EcoRI cuts 4 kb from one BamHI site and 8 kb from another BamHI site in the 12 Kb band produced by the BamHI digestion so the number of bands produced by the combined digest is 4.

size of the bands are 2kb,6kb, 4kb, and 8 kb.

Bam HI-2 kb, 6kb,12kb

EcoR I-20kb

EcoRI and BamHI- 2kb, 6kb, 4kb, and 8 kb.

2) Hpa I produce only one band so there is only one site for HpaI.

HindIII cuts pBLA230 to make 4 bands, so there are 4 HindIII sites, size of the bands are 13kb, 6kb, 4kb and 3kb. in the combined digest the bands obtained are 7 kb, 6 kb, 6kb, 4 kb and 3 kb so the HpaI makes a cut in the 13 kb band produced by the HindIII.

Hindi 6 kb 6 kb Hpal pBLA230 (26kb) Hindill 4kb 7 kb HindIII Hindill 3 kb