Puc57 vector

Manufactured by Tsingke

Sourced in China

The PUC57 vector is a plasmid commonly used in molecular biology experiments. It serves as a cloning vector, providing a means to insert and propagate DNA sequences of interest in bacterial host cells. The vector contains an origin of replication and a selectable antibiotic resistance marker, enabling the selection and maintenance of the recombinant plasmid in the host organism.

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Lab products found in correlation

Reverse transcriptase, by Vazyme (1 mentions) Exnase, by Vazyme (1 mentions) Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Nanodrop nd 2000, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Puc57

6 protocols using puc57 vector

Genotyping of HIV-1 Prevalent Strains in Guizhou, China

Cited 2 times

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In this study, 65 clinical serum specimens were collected from suspected HIV-infected patients from Guizhou Provincial Center for Clinical Laboratory between January 2023 and April 2023. Genomic RNA was isolated using Nucleic Acid Releasing Agent (GenDx Biotech Co., Ltd.) in accordance with the manufacturer’s instructions.
The full-length pol gene sequences of the representative genotypes of all known HIV-1 prevalent genotypes, including CRF01_AE, CRF07_BC, CRF08_BC, and subtype B, in China were downloaded from the GenBank database (genotype CRF01_AE: GenBank Accession No. U54771.1; genotype CRF07_BC: GenBank Accession No. U54771.1; genotype CRF08_BC: GenBank Accession No. AY008715.1; genotype subtype B: GenBank Accession No. GU647198.1) (He et al., 2012 (link); Zhao et al., 2018 (link)). The four genomic sequences were synthesized and cloned into the pUC57 vector by Tsingke Biotech (Beijing, China). The initial concentration of each plasmid was 1 × 10⁸ copies per milliliter, and the HIV-1 genotype CRF01_AE plasmid was used as a positive control.

Chen X., Du C., Zhao Q., Zhao Q., Wan Y., He J, & Yuan W. (2023). Rapid and visual identification of HIV-1 using reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticle-based lateral flow assay platform. Frontiers in Microbiology, 14, 1230533.

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SARS-CoV-2 N Protein Expression Constructs

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SARS-CoV-2 N protein-coding fragments were polymerase chain reaction (PCR)-amplified from the N protein expression construct in the PUC57 vector (Tsingke Biotech, Hangzhou, China) and subcloned into the pCS2 vector, while the coding region for SARS-CoV-2 N protein deletions (residues 1–175, residues 1–210, residues 51–C, residues 161–C, residues 211–C, and ΔSR) were amplified by PCR from a plasmid containing full-length SARS-CoV-2 N protein with appropriate sets of primers and was confirmed by sequencing. DNA fragments encoding human ubiquitin-specific peptidase 10 (USP10), double-stranded RNA-dependent protein kinase (PKR), and G3BP2 were PCR-amplified from cDNA that was reverse transcribed from HEK293T total RNA with reverse transcriptase (Vazyme, Nanjing, China) according to the manufacturer’s instructions. Exnase (Vazyme, Nanjing, China) was used to insert these sequences into the CMV-Flag or CMV-Myc backbone.

Luo L., Li Z., Zhao T., Ju X., Ma P., Jin B., Zhou Y., He S., Huang J., Xu X., Zou Y., Li P., Liang A., Liu J., Chi T., Huang X., Ding Q., Jin Z., Huang C, & Zhang Y. (2021). SARS-CoV-2 nucleocapsid protein phase separates with G3BPs to disassemble stress granules and facilitate viral production. Science Bulletin, 66(12), 1194-1204.

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Maintenance and Propagation of Cell Lines for ZIKV Research

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African green monkey kidney Vero cells (ATCC, CCL-81) and baby hamster kidney BHK-21 cells (ATCC, CCL-10) were maintained at 37 °C, 5% CO₂ in a high-glucose Dulbecco’s modified Eagle’s medium (DMEM) that was supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA, USA), 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and 1% Penicillin Streptomycin (P/S, Gibco, Waltham, MA, USA). The Aedes albopictus C6/36 cells (ATCC, CRL-1660) were maintained in RPMI 1640 Medium (Gibco, Waltham, MA, USA) supplemented with 10% FBS, 10 mM HEPES, and 1% P/S at 28 °C and 5% CO₂. ZIKV GZ01 strain (accession number KU820898) was kindly provided by Prof Cheng-Feng Qin from the Academy of Military Medical Sciences. Viral genomes and DNA linkers were de novo synthesized as DNA fragments and cloned into the pUC57 vector by Tsingke Biotech.

Dong H.L., He M.J., Wang Q.Y., Cui J.Z., Chen Z.L., Xiong X.H., Zhang L.C., Cheng H., Xiong G.Q., Hu A., Lu Y.Y., Cheng C.L., Meng Z.X., Zhu C., Zhao G., Liu G, & Chen H.P. (2023). Rapid Generation of Recombinant Flaviviruses Using Circular Polymerase Extension Reaction. Vaccines, 11(7), 1250.

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Monkeypox Virus Pseudovirus Production

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Full-length Congo Basin clade D14L (accession no. KP849471.1) and West African clade ATI (accession no. MT903346.1) sequences were synthetically produced and cloned into the pUC57 vector (Tsingke Biotech, China). The initial concentration of each plasmid was 1 × 10⁸ copies, and the two constructed plasmids acted as positive controls.
Two pseudoviruses (MPXV-D14L pseudovirus and MPXV-ATI pseudovirus) were constructed by Sangon Biotech Co., Ltd. (Shanghai, China), which were made with 293A cell cultures and included segments of the D14L gene (GenBank accession no. KP849471.1, genome coordinates 19294 to 19944) and ATI gene (GenBank accession no. MT903346.1, genome coordinates 135527 to 135918), respectively. Genomic DNA was extracted using viral qEx-DNA/RNA extraction kits (Xi′an Tianlong Science & Technology Co., Ltd., Xian, China) in accordance with the manufacturer’s instructions. The concentrations of genomic DNA were measured using a NanoDrop ND-2000 instrument (Thermo, USA) at A_260/280. Other microorganisms used in this study are shown in Table 1.

Chen X., Yuan W., Yang X., Shi Y., Zeng X., Huang J., Wang Y, & Li S. (2023). Ultrasensitive and Specific Identification of Monkeypox Virus Congo Basin and West African Strains Using a CRISPR/Cas12b-Based Platform. Microbiology Spectrum, 11(2), e04035-22.

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Comprehensive swine pathogen repository

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PCV2 (YHW strain) was preserved in our laboratory. The genomic DNA or cDNA of swine pathogens including porcine parvovirus (PPV, GD strain), classical swine fever virus (CSFV, shimen strain), Japanese encephalitis virus (JEV, sw/GD/2009 strain), porcine reproductive and respiratory syndrome virus (PRRSV, GD08-2 strain), pseudorabies virus (PRV, Ea strain), and Senecavirus A (SVA, GD-ZYY02-2018 strain) was also stored in our laboratory. DNA of African swine fever virus (ASFV) was a gift from other laboratory. PCV3 ORF2 gene was synthesized and cloned into the pUC57 vector by Tsingke Biological Technology (China). The synthetic plasmid was named as pUC57-PCV3 ORF2. Forty-two clinical tissue samples suspected to be PCV2 infection were collected from pig farms in Hebei province, China.

Chen W., Fan J., Li Z., Zhang Y., Qin Y., Wu K., Li X., Li Y., Fan S, & Zhao M. (2021). Development of Recombinase Aided Amplification Combined With Disposable Nucleic Acid Test Strip for Rapid Detection of Porcine Circovirus Type 2. Frontiers in Veterinary Science, 8, 676294.

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Quantification and Identification of C. trachomatis Genotypes

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A 456 bp fragment of the ompA gene was amplified from the clinical samples infected with C. trachomatis genotype B, D, E, F, G, H, J and K, and cloned into the pUC57 vector (TsingKe Biotech Corp, Beijing, China). The plasmid DNA was purified and quantified using a GENOVA NANO (Bibby Scientific Ltd., Stone, United Kingdom). The DNA copy number was calculated using the following formula: DNA copy number (copy number/μL) = [6.02 × 10²³ × plasmid concentration (ng/μL) × 10⁻⁹]/[DNA in length × 660]. A serial 10-fold diluted plasmid DNAs for C. trachomatis genotype B, D, E, F, G, H, J and K were used to determine the low detection limit of NGHTS. Furthermore, the mixtures of different plasmid DNAs of C. trachomatis genotype of F/G, E/J and E/F were prepared at the ratio of 50/50, 30/70, 20/80, 10/90, 2.5/97.5, 1/99, and amplified and sequenced to assess the sensitivity and accuracy of NGHTS in distinguishing mixed C. trachomatis genotypes.

Zhao J., Shui J., Luo L., Ao C., Lin H., Liang Y., Wang L., Wang H., Chen H, & Tang S. (2022). Identification and characterization of mixed infections of Chlamydia trachomatis via high-throughput sequencing. Frontiers in Microbiology, 13, 1041789.

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