Part:BBa_K1312004:Experience

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Applications of BBa_K1312004

This part is pSB1C3 which introduced 6xHis tag and stop codon UAA at the back of Biobrick Suffix and was made in order to add 6xHis tag to coding protein gene. If you add 6xHis tag to coding protein gene A, you need to get A gene from start codon to before stop codon by PCR.

This PCR product also need to have Biobrick Prefix and Pst1 recognision sight at the back of stop codon. If you can insert the PCR product, promoter and RBS to this vector, and transforme the plasmid into E.coli, you can get protein conjugated to 6xHis tag.

User Reviews

We introduce some protein coding genes into BBa_K1312004 and generate protein-6xHistag conjugates. Then, we conduct western blotting using 6xHistag antibody which fluorescent labeled by HRP.

Fig. 1

This result shows that 6xHistag added to the protein from REDOX gene by BBa_K1312004, because mokecular weight of REDOX-6xHistag conjugate is 27.25kDa and main band is around 25kDa.About the protein from SAMmt gene and 6xHistag conjugate, molecular weight is 48.53kDa, the band of this conjugate shown around 50kDa. Therefore 6xHistag also added to the proteins.

UNIQ46c55452ef6b58e5-partinfo-00000002-QINU UNIQ46c55452ef6b58e5-partinfo-00000003-QINU