Difference between revisions of "Part:BBa K4099000"

Latest revision as of 11:04, 20 October 2021

sgRNA

Profile

Name: sgRNA

Base Pairs: 112bp

Origin: Lactobacillus casei, genome

Properties: A piece of RNA

Usage and Biology

BBa_K4099000 is a piece of RNAs that function as guides for RNA- or DNA-targeting enzymes, which they form complexes with.

Figure1. Principle diagram of CRISPR-Cas9..

The sgRNA and HR are inserted in the pLCNICK vector.

Proof of function

We electrotransformed the plasmid pIB165 as the foreign DNA to test the transformation efficiency of our modified L. casei, it took several days to culture and finally saw the comparison results. As showing above, we can see that the modified L. casei (KO) has higher transformation efficiency with remarkably more colonies than the wild (Wild).

1.Transformation test result

Figure 2. Comparison between the wild L. casei (left) and modified L. casei (right, LSEI-2094 knocked out) in transformatio..

Graph 3. Comparison between the wild L. casei and modified L. casei in transformation

Figure 3. Histogram comparison between wild strain and KO strain..

In addition, we measured OD600 of these strain groups which were pre-spread plates with different volumes of bacteria solutions so as to quantify the transformation results as showing above (Fig. 3). In conclusion, it indicates that our modified L. casei has much higher efficiency of the foreign plasmid transformation than the wild and the modified L. casei has great potential to be used as the recombinant carrier in various areas.

Sequence and Features