Synthetic gene blocks

Manufactured by Integrated DNA Technologies

Synthetic gene blocks are DNA fragments that are artificially synthesized. They are designed to serve as modular components for constructing larger, more complex genetic sequences. These gene blocks can be used as building blocks in various molecular biology applications.

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Gene blocks

2 protocols using synthetic gene blocks

Molecular Biology Techniques Protocol

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All enzymes used for molecular biology were from New England Biolabs. Synthetic oligonucleotides for cloning and sequencing were purchased from Eurofins Genomics, and synthetic gene blocks were purchased from Integrated DNA Technologies. Epoch Life Science kits were used for plasmid DNA purification. Sources of noncanonical amino acids are listed below in the “Yeast transformations, propagation, and induction” section.

Stieglitz J.T., Kehoe H.P., Lei M, & Van Deventer J.A. (2018). A robust and quantitative reporter system to evaluate noncanonical amino acid incorporation in yeast. ACS synthetic biology, 7(9), 2256-2269.

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Activin A Mutein Production in CHO-K1 Cells

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The mature regions of Activin A or Activin A muteins were cloned downstream of mouse Inhba pro-peptide using isothermic Gibson assembly (Gibson et al., 2010 (link)) of synthetic gene blocks (Integrated DNA Technologies) into a CMV-based mammalian expression vector. The corresponding expression vectors were introduced into CHO-K1 cells expressing human Furin (CHO-KI.Furin). The resulting conditioned media was concentrated approximately ten-fold using a filtration column with a 3K molecular weight cut off (Pierce) before biochemical characterization of the Activin A muteins in cell-based assays. Following this primary analysis, Activin A and the Activin A F2TL mutants were stably expressed in CHO-K1.Furin cells. The Activin A pro-peptide complex was first purified to homogeneity by heparin chromatography followed by size exclusion chromatography on a preparative scale S200 column. To isolate the mature Activin A mutein dimers, reverse phase chromatography on C4 column was used, and mature Activin A was eluted with an acetonitrile gradient. The sequence information of Activin A muteins is available on Dryad (doi:10.5061/dryad.5hqbzkh3d).

Aykul S., Corpina R.A., Goebel E.J., Cunanan C.J., Dimitriou A., Kim H.J., Zhang Q., Rafique A., Leidich R., Wang X., McClain J., Jimenez J., Nannuru K.C., Rothman N.J., Lees-Shepard J.B., Martinez-Hackert E., Murphy A.J., Thompson T.B., Economides A.N, & Idone V. (2020). Activin A forms a non-signaling complex with ACVR1 and type II Activin/BMP receptors via its finger 2 tip loop. eLife, 9, e54582.

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