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Correction to: Autolysis of Pichia pastoris induced by cold

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AMB Express20188:185

https://doi.org/10.1186/s13568-018-0712-2

  • Received: 2 November 2018
  • Accepted: 2 November 2018
  • Published:

The original article was published in AMB Express 2017 7:95

Correction to: AMB Expr (2017) 7:95 https://doi.org/10.1186/s13568-017-0397-y

The original version of this article (Bartolo-Aguilar et al. 2017) was written and published including the first construction strategy of pLGC09, but not the final one. This error was pointed out by a reader and an analysis of sequences of parts of the plasmid corroborated this. The final construction strategy was reanalysed and confirmed the error. This error affected the text, Table 2, Fig. 1 and the Additional files, but did not affect the results and conclusions stated in the paper. The authors regret that this error occurred in the original publication of the article. The corrected text, Table 2 and Fig. 1, and the Additional files (Additional file 1. Construction strategy of pLGC09 and Additional file 2. Plasmid pLGC09) are given in this correction.
Table 2

Primers used in this study

Primera

Sequence (5′→3′)b

Restriction enzyme

Underlined sequencec

YL1-F

GGGGtcgcgaAATCTTCCTCAGAAGAGGAAT

NruI

Homologous Pcctα region corresponding to nucleotides (nt) 886843 to 886863

YL1-R

GCGCggatccTGAATAATTGGGACATCGCTAA

BamHI

Homologous Pcctα region corresponding to nt 887226 to 887247

YL2-F

CCCCagatctTTCTCTCGTTAGCTTCCCAAA

BglII

Homologous eng region corresponding to nt 802080 to 802100

YL2-R

GCGCctcgagGTCAACTATGTGTGGTACCTT

XhoI

Homologous eng region corresponding to nt 805196 to 805216

YL3-F

GCGCgtcgacTGAGTTTTAGCCTTAGACATGACT

SalI

Homologous aox1 TT region corresponding to nt 890 to 913

YL3-R

GGGGatttaaatGGGGATCCGCACAAACGAA

SwaI

Homologous aox1 TT region corresponding to nt 1217 to 1235

YL4-F

CCCCatttaaatGGATCCCCCACACACCATA

SwaI

Homologous Ptef1 region corresponding to nt 965 to 983

YL4-R

GCGCcacgtgCACATGTTGGTCTCCAGCTT

PmlI

Homologous cyc1 region corresponding to nt 2157 to 2138

YL5-F

GGGGgccggcCCAAGTCCAAGGTCTCCAAT

NaeI

Homologous leu2 region corresponding to nt 86424 to 86443

YL5-R

GGGGcacgtgTTTCCGAACCTTACAGTAGAG

PmlI

Homologous leu2 region corresponding to nt 87940 to 87920

aF forward primer, R reverse primer

bLowercase letters indicates the restriction endonuclease recognized site denoted in the next column

cNucleotide numbering has been taken according to the sequence depicted in the access number mentioned in the section of Materials and methods

Fig. 1
Fig. 1

Structure of pLGC09. Plasmid pLGC09 comprises three modules: a the lytic regulated module is composed of the promoter of chaperonin CCT one complex (Pcctα) that drives expression of eng in P. pastoris and it is finished by aox1 transcription termination (aox1 TT) region; b the selection marker module is composed of the translational elongation factor 1 gene promoter (Ptef1) and of the EM7 synthetic prokaryotic promoter (PEM7) that drive expression of the ble gene in P. pastoris and E. coli respectively, these expressions are finished by cyc1 transcription termination region (cyc1 TT); c the homologous recombination module is composed of the leu2 functional gene including its promoter and transcription termination region; and d the replicon pCR®4Blunt-TOPO® that includes the functional gene bla and pUC origin (oripUC)

The second paragraph in the section Materials and methods entitled, “Construction of plasmid pLGC09” was wrong and should read:

“Three modules were obtained: First, to construct the lytic module, the promoter Pcctα amplified from the genomic DNA of S. cerevisiae (amplicon YL1; Table 2 shows the corresponding primers used to amplify each amplicon, i.e. YL1-F and YL1-R to amplify amplicon YL1) was cloned in pCR®4Blunt-TOPO® (using the Zero Blunt® TOPO® PCR Cloning Kit for Sequencing (Invitrogen) following the instructions of the supplier; pLGC01), the gene eng amplified from the genomic DNA of P. pastoris GS115 (amplicon YL2) was cloned in PmeI digested pLGC01 using T4 DNA Ligase to obtain pLGC03. This plasmid digested with XhoI and aox1 TT amplified from pGAPZαA (Invitrogen; amplicon YL3) digested with SalI were ligated together (Table 2 shows the restriction sites used to construct the module, the joining of XhoI to SalI digested ends are cohesive but their joining does not let the restriction sites to be regenerated), and amplified using primers YL1-F and YL3-R (amplicon YL1-YL3; lytic module). The amplicon was cloned in pCR®4Blunt-TOPO® (pLGC05). Second, the selection marker module consisted of a fragment containing the promoters Ptef1 and PEM7, ble, and the cyc1 TT was directly amplified from pGAPZαA (amplicon YL4). Third, the full-length leu2 gene including its promoter and transcription terminator was amplified from the genomic DNA of P. pastoris (amplicon YL5) to be the integration module. The last two modules were cloned each in pJET1.2/blunt using the Clone-JET™ PCR Cloning Kit (Fermentas; Waltham, MA, USA; pLGC06 and pLGC07, respectively) according to the instructions of the manufacturer. The plasmid containing the selection marker module was digested with PmlI, ligated to the plasmid including the integrative module digested with NaeI (both enzymes produce blunt ends), and amplified through PCR using the primers YL4-F and YL5-R. The two-modules amplicon was cloned in pCR®4Blunt-TOPO® (pLGC08). The final construction was assembled by subcloning the fragment NotI–PmlI from the plasmid including the selection marker and integration modules in NotI–NruI digested pCR®4Blunt-TOPO®-lytic module using classical restriction and ligation techniques (Sambrook and Russell 2001), obtaining at last the vector pLGC09 (details on the construction strategy, and pLGC09 data could be consulted in Additional files 1, 2).”

Table 2 was corrected, including the sequences of the primers used to construct pLGC09.

These corrections do not alter any conclusions or results of our study.

The authors wish to apologize for this confusion and the inconvenience that this might have caused.

Notes

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Authors’ Affiliations

(1)
Department of Biotechnology and Bioengineering, Cinvestav-IPN, Av. Instituto Politécnico Nacional 2508, Col. San Pedro Zacatenco, 07360 Gustavo A. Madero, CDMX, Mexico
(2)
Department of Microbiology, Escuela Nacional de Ciencias Biológicas del IPN, Prol. Carpio y Plan de Ayala S/N Col. Santo Tomás, 11340 Miguel Hidalgo, CDMX, Mexico

Reference

  1. Bartolo-Aguilar Y, Dendooven L, Chávez-Cabrera C, Flores-Cotera LB, Hidalgo-Lara ME, Villa Tanaca L, Marsch R (2017) Autolysis of Pichia pastoris induced by cold. AMB Expr 7:95. https://doi.org/10.1186/s13568-017-0397-y View ArticleGoogle Scholar

Copyright

© The Author(s) 2018

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