The following is the partial sequence of a bacterial gene ORF: 5’ --------------------CGGAATTCCCGGGGATCC------------------------3’ (The remaining sequence of the gene ORF never affects the cloning of this gene. The multiple cloning sites of the plasmid vector can be cleaved by 8 restriction endonucleases that are listed in the table.5’- and 3’-end sequences of the bacterial gene ORF are 5’ ATGGAGT TATCCAGGTGCCT--- and 3’AATATGGAGTTATCCAGGTGCCT---, respectively. After choosing the appropriate restriction endonuclease(s) for directional cloning using the table below, determine the sequences of two 20-mer primers used for the PCR amplification of the gene ORF. Table 18.2 Characteristics of some common type II restriction enzymes used in recombinantDNA technologyMicroorganism fromType of fragmentEnzymewhich enzyme is isolatedRecognition sequenceend producedBamHIBacillus amyloliquefaciens5-GGATCC-3Cohesive3'-CCTAGG-3'EcoRIEscherichia coli5'-GAATTC-3Cohesive3'-CTTAAG-5'EcoRIIEscherichia coli5-CCAGG-3'3'-GGTCC-5'CohesiveHaelllHaemophilus aegyptius5-GGCC-3'3-CCGG-5BluntHindIIIHaemophilus influenzae5'-AAGCTT-3'3'-TTCGAA-5CohesiveNotlNocardia otitidis-caviarum5-GCGGCCGC-3Cohesive3'-CGCCGGCG-5PstlProvidencia stuartii5-CTGCAG-3'Cohesive3'-GACGTC-5'↑SmalSerratia marcescens5-CCCGGG-3'Blunt3'-GGGCCC-5'

Primers for PCR amplification are designed by integrating restriction sites for directed cloning and…

Answered: Table 18.2 Characteristics of some…

Biology Today and Tomorrow without Physiology (MindTap Course List)

5th Edition

ISBN:9781305117396

Author:Cecie Starr, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Christine Evers, Lisa Starr

Chapter10: Biotechnology

Section: Chapter Questions

Problem 8SQ

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The following is the partial sequence of a bacterial gene ORF: 5’ --------------------CGGAATTCCCGGGGATCC------------------------3’ (The remaining sequence of the gene ORF never affects the cloning of this gene. The multiple cloning sites of the plasmid vector can be cleaved by 8 restriction endonucleases that are listed in the table.

5’- and 3’-end sequences of the bacterial gene ORF are 5’ ATGGAGT TATCCAGGTGCCT--- and 3’AATATGGAGTTATCCAGGTGCCT---, respectively. After choosing the appropriate restriction endonuclease(s) for directional cloning using the table below, determine the sequences of two 20-mer primers used for the PCR amplification of the gene ORF.