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Ptrchis2 vector

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The PTrcHis2 vector is a plasmid used for the expression and purification of recombinant proteins in Escherichia coli. It contains a trc promoter, a polyhistidine (His) tag sequence, and other elements necessary for protein expression and purification.

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Lab products found in correlation

Ncoi hindiii digested ptrchis2 vector, by Thermo Fisher Scientific (2 mentions) B per, by Thermo Fisher Scientific (1 mentions) E coli strain nico21 de3, by New England Biolabs (1 mentions) Ni nta column, by Qiagen (1 mentions)

Spelling variants of this product

NcoI/HindIII digested pTrcHis2 vector (4 citations)

3 protocols using ptrchis2 vector

1

Heterologous Expression of Acyl-CoA Synthases

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Example 16

Homologs to E. coli fadD can be expressed in E. coli by synthesizing codon-optimized genes based on a desired sequence from M. tuberculosis HR7Rv (NP_217021, FadDD35), B. subtilis (NP_388908, YhfL), Saccharomyces cerevisiae (NP_012257, Faa3p) or P. aeruginosa PAO1 (NP_251989). The synthetic genes can be designed to include NcoI- and HindII-compatible overhangs. The acyl-CoA synthases can then be cloned into a NcoI/HindIII digested pTrcHis2 vector (Invitrogen Corp., Carlsbad, Calif.) as described above and expressed in E. coli strain MG1655 ΔfadE. The expression in E. coli may lead to an increased production of acyl-CoA.

Fatty acid derivatives such as an FAEE can also be produced by co-transformation of the E. coli strain MG1655 ΔfadE with various acyl-CoA synthases in the pTrcHis2 vector with a compatible plasmid derived from pCL1920, which contains the thioester gene from Cuphea hookeriana with or without an ester synthase from A. baylyi. The resulting production host will produce FAEE when cultured in a medium containing ethanol as described above.

US11021695B2. Methods and compositions related to thioesterase enzymes (2021-06-01). GENOMATICA, INC. [US]. Inventors: Louis Hom [US], Na Trinh [US], Murtaza Alibhai [US], Zhihao Hu [US], Eli Groban [US], Vikranth Arlagadda [US], Elizabeth Clarke [US].
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2

Heterologous Expression of Acyl-CoA Synthases

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 16

Homologs to E. coli fadD can be expressed in E. coli by synthesizing codon-optimized genes based on a desired sequence from M. tuberculosis HR7Rv (NP_217021, FadDD35), B. subtilis (NP_388908, YhfL), Saccharomyces cerevisiae (NP_012257, Faa3p) or P. aeruginosa PAO1 (NP_251989). The synthetic genes can be designed to include NcoI- and HindII-compatible overhangs. The acyl-CoA synthases can then be cloned into a NcoI/HindIII digested pTrcHis2 vector (Invitrogen Corp., Carlsbad, Calif.) as described above and expressed in E. coli strain MG1655 ΔfadE. The expression in E. coli may lead to an increased production of acyl-CoA.

Fatty acid derivatives such as an FAEE can also be produced by co-transformation of the E. coli strain MG1655 ΔfadE with various acyl-CoA synthases in the pTrcHis2 vector with a compatible plasmid derived from pCL1920, which contains the thioester gene from Cuphea hookeriana with or without an ester synthase from A. baylyi. The resulting production host will produce FAEE when cultured in a medium containing ethanol as described above.

US09951322B2. Methods and compositions related to thioesterase enzymes (2018-04-24). REG Life Sciences, LLC [US]. Inventors: Louis Hom [US], Na Trinh [US], Murtaza Alibhai [US], Zhihao Hu [US], Eli Groban [US], Vikranth Arlagadda [US], Elizabeth Clarke [US].
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3

Cloning and Purification of Murine Chromogranin A

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Full length murine CHgA was amplified from the pCMV6-Kan/Neo cDNA clone (Origene) with the following primers: F, 5’-TCTAGATCTTTCCGCACCGTCCG-3’, 5’-GTCAGAATTCCCTACTCGAGCAGCAGTC-3’. The cycling parameters were as follows: 94°C for 6 mins, followed by 39 cycles of 94°C for 30 sec, 56°C for 60 sec, 72°C for 60 sec, and 72°C for 10 mins. The PCR product was ligated into the pTrcHis2 vector (Invitrogen). The pTrcHis2 vector was transformed into E. coli strain NiCo21 (DE3) (New England BioLabs) and the expression of HIS-tagged proteins was induced with isopropyl β-D-1-thiogalactopyranoside (IPTG) at 37°C for 3.5 hr. The bacteria were lysed using B-PER (Thermo Scientific), following the manufacturer’s instructions. CHgA was purified using the NI-NTA column (Qiagen), following the manufacturer’s instructions.
Marré M.L., Profozich J.L., Coneybeer J.T., Geng X., Bertera S., Ford M.J., Trucco M, & Piganelli J.D. (2016). Inherent ER stress in Pancreatic Islet β cells Causes Self-Recognition by Autoreactive T Cells in Type 1 Diabetes. Journal of autoimmunity, 72, 33-46.
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