(0) Draft Berat (301) II Ndet (302) Snabt (68) VPLS (forward primer) (09.856) Ec
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(0) Draft Berat (301) II Ndet (302) Snabt (68) VPLS (forward primer) (09.856) Eco53k (924) EcoRI BamHI XbAI Mrel Mlul GOIN Hindl|I Notl HinPIL EcoRI (1979 BamHI (982) Acc6ST (9) Kont (1002) -Asist - Soft (1024) Miret - SgrAI (1026) Aset - BHEI (100) Hindtit (04) Rsrt (1061) Miut (1967) Net (2017) PaeR21 - PapxI - Tichot (1095) Panel (1094) PCMV6-A-GFP HGH pognal Alet (114) XL39 (reverse primer) (1200 1219) Obst (317) BibvCt-Boulot (1420) Bomb! (1484) DatXI (1571) Agel (1682) (3979) ANI SAI. LBS) Svo povA) signal OS) Peit -SHI 2955) Stut (201) Avril (2102) Bcle (2153) Beiwt (2366) Kast (2322) Nar! 2023 Stel (324) PluTI (2326) Aart (2626 (31) BetE171 BsaBI (2638) O2 Bemt PspOMI (2799) Apal (2763) (2987) Plot (2894) BstBPstr (2878)Explanation / Answer
A promoter is a cis element that shows directionality. So, we have to clone the ATG of the GOI facing the 3'-end of the promoter.
Assumption: ATG of GOI is towards the side of EcoRI in vector A.
We can not use restriction sites that located inside the gene coding sequence as they would lead to the production of truncated peptides.
Enzyme sites in Vector A
5' possible enzymes = Eco RI, Bam HI and Xba I
3' possible enzymes = Hind III, Not I and Hin PII
Enzyme sites in Vector B
5' possible enzymes: Eco RI and Bam HI
3' possible enzymes = Hin DIII and Not I
Possible enzyme combinations that can be used to clone GOI into vector B:
i. Eco RI and Hind III
ii. Eco RI and Not I
iii. Bam HI and Hind III
iv. Bam HI and Not I
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