Difference between revisions of "Part:BBa K4335013"

Revision as of 07:17, 11 October 2022 (view source) ZXFeng (Talk | contribs) ← Older edit		Revision as of 07:28, 11 October 2022 (view source) ZXFeng (Talk | contribs) Newer edit →
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	<h2>Result</h2>		<h2>Result</h2>
	<h3>Plasmid Construction</h3>		<h3>Plasmid Construction</h3>
−		+	We used the primer with a length of about 600 bp designed according to the Hyg resistance gene to carry out PCR validation on the single colonies successfully transferred into pTX2038 and pTX2040.
	<figure>		<figure>
	<img src="https://static.igem.wiki/teams/4335/wiki/zhjt2.jpg" width="50%" style="float:center">		<img src="https://static.igem.wiki/teams/4335/wiki/zhjt2.jpg" width="50%" style="float:center">
	<figcaption>		<figcaption>
−	<p style="font-size:1rem">	+	<p style="font-size:1rem">Single algae drop amplifies the Cas9 gene.
	</p>		</p>
	</figcaption>		</figcaption>

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Revision as of 07:28, 11 October 2022

SpCas9

The Cas9 protein is part of the bacteria's CRISPR/Cas9 immune defense mechanism to identify and destroy foreign DNA. By incorporating the foreign DNA into the bacteria's own DNA, it has a memory of any prior foreign DNA that the bacteria has encountered.

The Cas9 endonuclease can be used either with the tracrRNA and crRNA (CRISPR RNA- Clustered Regularly Interspaced Short Palindromic Repeats) or with a guide RNA to cleave foreign DNA at a species locations. This makes exceptionally useful for DNA cloning in synthetic biology.

Usage

We found a SpCas9 from iGEM16_Cambridge-JIC in the previous part [BBa_K2148013] .

This part is based on the original Cas9 part in the registry, BBa_K1218011. To enhance the expression of Cas9 in the chloroplast chassis, Cambridge-JIC have codon-optimised it using the codon-optimisation software developed by Saul Purton.

The amino acid and nucleotide sequences of BBa_K2148013 and BBa_K2148013 were compared.

Result

Plasmid Construction

We used the primer with a length of about 600 bp designed according to the Hyg resistance gene to carry out PCR validation on the single colonies successfully transferred into pTX2038 and pTX2040.

Functional Identification

See the part [BBa_K4335020] for result.

Sequence and Features