Team:BioIQS-Barcelona/Parts

BIO IQS

Basic Parts

Our basic parts consist of the alpha and beta chains of the HLA-DQ protein. Both chains can be obtained from genomic DNA of any celiac patient by PCR following the protocol that we designed in our basic parts page.

However, to get and submit the parts, we synthetized them after performing a codon optimization of the sequence for expression in E. coli. The aminoacidic sequence was obtained from the genomic sequence of a celiac patient. The parts were then amplified by PCR and cloned into the pSB1C3 vector.

BBa_K2879666: HLA-DQA:

This biobrick corresponds to the HLA-DQA1*05:01 gene variant. Thus, when coupled to the protein encoded by the HLA-DQB1*02:01 gene variant (BBa_K2879800) it will form the HLA haplotype DQ2. This part only contains the exons 2 and 3 of the protein (which encode the extracellular domains of the protein), for more information see the design tab. Therefore, this part (HLA-DQA1) can be used in combination with part BBa_K2879800 (HLA-DQB) to form a variant of the HLA-DQ complex which recognizes the deaminated gliadin peptide, which plays a central role in celiac disease.

Note that this biobrick has been synthetized de novo and it has been codon optimized to be expressed in E. coli BL21. See the detailed information of this biobrick for more information on how to form the complex HLA-DQ by expressing this part in combination with part BBa_K2879800 in E. coli. See the design tab to see how the sequence has been obtained and codon optimized.

BBa_K2879800: HLA-DQB:

This biobrick corresponds to the HLA-DQB1*02:01 gene variant. Thus, when coupled to the protein encoded by the HLA-DQA1*05:01 gene variant (BBa_K2879666) it will form the HLA haplotype DQ2. This part only contains the exons 2 and 3 of the protein (which encode the extracellular domains of the protein), for more information see the design tab. Therefore, this part (HLA-DQB1) can be used in combination with part BBa_K2879666 (HLA-DQA1) to form a variant of the HLA-DQ complex which recognizes the deaminated gliadin peptide, which plays a central role in celiac disease.

Note that this biobrick has been synthetized de novo and it has been codon optimized to be expressed in E. coli BL21. See the detailed information of this biobrick for more information on how to form the complex HLA-DQ by expressing this part in combination with part BBa_K2879666 in E. coli. See the design tab to see how the sequence has been obtained and codon optimized.

Composite Parts

We have created several functional units made from the combination of the HLA-DQ basic parts. The objective of our composite parts is to find a suitable expression system for the HLA-DQ protein, which is the basis of our sensor.

BBa_K2879100: HLA-DQ a+b polycistronic

This composite is formed by the α and β chain of the human HLA-DQ flanked by the restriction enzymes NdeI and SacI/SalI. Each chain is separated by an intergenic regions containing a RBS for translation.

BBa_K2879102: LZipper+HLADQA+HLADQB+NdeISacI

This composite encodes for the α and β chains of the human HLA-DQ. Each chain is linked to a Fos/Jun domain for protein stability. Both chains are separated by an intergenic region that contains a RBS. The entire construct is flanked by the restriction enzymes NdeI and SacI/SalI. A Strep Tag is placed after the α chain.

Submitted parts

Below you can see a table of all the parts that our team has characterized and submitted to the registry.

<groupparts>iGEM2018 BioIQS-Barcelona</groupparts>