Recombinant plasmid was transformed to E. coli BL21 and cultured at 37 °C for 12 h. positive monoclonal bacteria were cultured and verified by PCR.
Figure1: 1% Agarose Gel Electrophoresis of PCR, which shows that our vector was successfully constructed.
In order to verify whether the cell expression of Lysin can perforate on the plasma membrane, we test the growth curve of recombinant E.coli and check them under the scanning electron microscope. Recombinant bacteria is cultured by LB medium adding with 0.1% kanamycin till logarithmic phase and induced by IPTG. Then culture solution is transferred into 96-well microtiter plates and measured the light absorption of 600 nm.
Figure2 Cell growth curve with or without IPTG induction. E. coli without IPTG induction grew normally while the density of E.coil decreased afte addition of IPTG.
Figure3 The images are from scanning electron microscopes.(a)Before induction.(b)After induction.There are holes in the plasma membrane after induction, which can prove that lysin expresses successfully.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC