Sac 1

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Sac I is a laboratory equipment product designed for DNA restriction enzyme digestion. It is a type II restriction endonuclease that recognizes and cleaves the DNA sequence 'GAGCTC'.

Automatically generated - may contain errors

Lab products found in correlation

Ppicza, by Thermo Fisher Scientific (1 mentions) P pastoris strain gs115, by Thermo Fisher Scientific (1 mentions) E coli dh5α, by Takara Bio (1 mentions) Ecori, by Thermo Fisher Scientific (1 mentions) Xhoi, by Thermo Fisher Scientific (1 mentions) Ppic9k, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Sac I restriction enzyme (2 citations)

2 protocols using sac 1

Efficient Construction and Validation of HOG1 Gene Knockout in Cryptococcus gattii

Cited 2 times

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Using restriction enzyme identification, PCR and sequence analysis (25 (link)), the recombinant plasmid pGAPza-HOG1 containing the intact HOG1 gene was successfully constructed. The HOG1 gene knockout expression vector pGAPza-dHOG1 was constructed by knocking out 400-bp fragment of pGAPza-HOG1 using a single restriction, Sac I (Invitrogen, Thermo Fisher Scientific, Inc.). The knockout product was transformed into Cryptococcus gattii CZ2012 cells by electroporation (electric shock condition: 1,500 V, 400 Ω, 25 µF, 5 mS; twice, internal: 5 min). Stable transformants were obtained through screening on YPD medium containing zeocin and subsequently confirmed by diagnostic PCR, DNA sequencing and Southern blot analysis (25 (link)).
To construct the hog1Δ+HOG1 reconstituted strain, pGAPza-HOG1 was linearized and transformed into the mutant (deletion of HOG1) by electroporation (26 (link)). Stably transfected colonies were selected on medium containing ampicillin and zeocin. The reconstitution of the HOG1 gene was confirmed by diagnostic PCR and Southern blotting (25 (link)).

Huang Y.M., Tao X.H., Xu D.F., Yu Y., Teng Y., Xie W.Q, & Fan Y.B. (2021). HOG1 has an essential role in the stress response, virulence and pathogenicity of Cryptococcus gattii. Experimental and Therapeutic Medicine, 21(5), 476.

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Heterologous Protein Expression in P. pastoris

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The strains, plasmids, and primers used in this study are listed in Table 1. E. coli DH5α, purchased from Takara (Dalian, China), was used for plasmid proliferation. P. pastoris strain GS115, two yeast expression vectors, pPIC9K and pPICZA, as well as all restriction enzymes (EcoRI, NotI, XhoI, SalI, SacI, and PmeI) were purchased from Invitrogen (Carlsbad, CA, USA).
Luria-Bertani (LB) medium contained 10 g/L sodium chloride, 10 g/L tryptone, and 5 g/L yeast extract. The yeast extract peptone dextrose (YPD) medium contained 20 g/L glucose, 20 g/L tryptone, and 10 g/L yeast extract. The minimal dextrose (MD) medium contained 13.4 g/L yeast nitrogen base (YNB), 4 × 10⁻⁴ g/L biotin, and 20 g/L glucose. The buffered glycerol-complex (BMGY) medium contained 20 g/L tryptone, 10 g/L yeast extract, 13.4 g/L yeast nitrogen base (YNB), 4 × 10⁻⁴ g/L biotin, 100 mmol/L potassium phosphate buffer (pH 6.0), and 10 g/L glycerol. Buffered methanol-complex (BMMY) medium contained 20 g/L tryptone, 10 g/L yeast extract, 13.4 g/L yeast nitrogen base (YNB), 4 × 10⁻⁴ g/L biotin, 100 mmol/L potassium phosphate buffer (pH 6.0), and 5 g/L methanol.

Jiang L., Guan X., Liu H., Chang X., Sun J., Sun C, & Zhao C. (2023). Improved Production of Recombinant Carboxylesterase FumDM by Co-Expressing Molecular Chaperones in Pichia pastoris. Toxins, 15(2), 156.

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