Solution 2/3

A. Digest vector pQE-60 with BamHI [G^GATCC] (121) and HindIII [A^AGCTT] (152).

5'--G.GGA.G/GA.TC C.AGA.TCT.CAT.CA-.---CAC.TA/A.GCT. TAA.TTA.GCT.GAG--3'
3'--C.CCT.C CT.AG/G.TCT.AGA.GTA.GT-.---GTG.AT T.CGA./ATT.AAT.CGA.CTC--5'

Resulting fragments following this pQE-60 digestion:

1. 3400 bp HindIII 152 - BamHI 121
2.    31 bp BamHI 121 - HindIII 152

B. Digest insert pGEM-GOI with BamHI [G^GATCC] (2915) and HindIII [A^AGCTT] (3266).

5'--C.TCC.G/GA.TC C.ATG.GTT.CTT.CC-.---.TAG.A/AG.CT T.ATC.GAT.TTC.GA--3'
3'--G.AGG.C CT.AG/G.TAC.CAA.GAA.GG-.---.ATC.T TC.GA/A.TAG.CTA.AAG.CT--5'

Resulting fragments following this pGEM-GOI digestion:

1. 2991 bp HindIII 3266 - BamHI 2915
2.   351 bp BamHI 2915 - HindIII 3266

Let's examine this solution in more detail. Look at the table below:


Insert
pGEM-GOI
Vector
pQE60

The resulting plasmid will have the following features:
 
Solution number
pQE60-GOI will contain
pQE60-GOI will NOT contain
1
BamHI & HindIII
 Ori
2
Two sites of EcoRI
 Lac Promotor
3
 Amp Resistance
Stop codon
4
 Four additional amino acids upstream to GOI
 PolyHis