Xmal Zral BsrDI 4234 Acll 4160 not available from NEB BsmBI

Xmal, Zral BsrDI 4234 Acll 4160 not available from NEB BsmBI 4028 Dral 3989 Mscl 3 98 Nicol Styl 3945 Sspl 3936 EcoRI Scal 383 BspEI 5 Bme 15801 3779 Dral 83 BstBI 3717 Pvull 103 PflFI Tth1111 3699 BsaAI 310 Drdl 3696 BsmBI 324 Btsl 36200 Bsu36I 3592 Pvull 515 Bell 3542 Bmtl hel 583 BstZ17I Accl 594 BsaBI 3442 635 BstYI 344 Xmnl 665 BspEI 3438 Bsgl Bsgl 3424 SgrAI 668 PshAI 330 BsrBI 755 Bogl 3297 SacII 832 BsrBI 3223 BssSI 9600 HincII 3212 pACYC184 Ahdl 3204 Acul 3151 4,245 bp Nispl 1158 BsaAI 3142 PpuMI 2975 BseRI Mscl 2939 BseRI 1269 PpuMI 2933 BstBI 1319 Aval BsoBI 2920 1406 Asel Styl 2864 Sspl 1419 2663 NmeATII Xbal 1425 Bgll 2658 Earl 1447 BspMI BfuAl 2558 Acul 1464 BstAPI 2540 BspDI cial 1518 Nrul 2467 Tc HindIII 1524 BseYI 2444 NmeAIII 1611 Eagl 2434 Sfcl 633 Bgll 2424 Acll 2395 EcoRV 1680 Brntl -Nhel 1724 PshAI 2207 BamHI BstYT 1870 Bcgl 2203 SgrAI 1904 Sall Accl HinclI 2146 Banll 1966 EcoNI 2117 Banll 1980 Sphl Nspl 2057 BspHI 1984

Solution

pETBlue-2 is used for screening the clones.

The lacI gene comes from pACYC184. Successful clones having disrupted LacI will show white color, while clones without LacI disruption will give a blue color, which can be ruled out.

chloramphenicol is used as a marker because of its property of translation inhibition. pACYC184 is an expression vector, and protein expression can be checked by using chloramphenicol resistance behavior of the clone. While ampicillin\'s nature is to interfere with host cell wall formation. thus chloramphenicol resistance gene is important for expression strain.