3.8  Codon Optimization and In Silico Cloning
In the follow up experimental studies, the maximum expression of MEPVC is highly desirable [35]. One requirement for that is the codon usage of MEPVC that must be adapted according to the expression system, for instance, here we used E. coli K12 as a MEPVC expression system. The codon adaptation index (CAI) and GC content revealed for the improved sequence are highly satisfactory with value of 0.96 and 48.85, respectively strongly indicating high MEPVC expression. The MEPVC then enclosed on both sites by 6x histidine tag to ease its purification process and inserted at appropriate sites of pET28a(+) vector as shown in Figure 6 .
Fig. 6 In silico restriction cloning of the MEPVC into pET28a(+) vector. The insert is shown in red.