首页 » Nisin诱导调控乳酸菌表达载体系统及宿主菌-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

Nisin诱导调控乳酸菌表达载体系统及宿主菌-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

  • 价  格:¥17930
  • 货  号:Nisin Controlled Gene Expression System for L. lactis
  • 产  地:北京
点击询问我要采购
 竭诚为您服务!
BioVector NTCC典型培养物保藏中心
联系人:Dr.Xu, Biovector NTCC Inc.

电话:400-800-2947 工作QQ:1843439339 (微信同号)

邮件:Biovector@163.com

手机:18901268599

地址:北京

已注册
 

The Effective Nisin Controlled Gene Expression System for L.
lactis

Nisin诱导调控乳酸菌表达载体系统-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心




Controlled gene expression in L. lactis – an emerging alternative
to recombinant protein production



Small scale L. lactis culture



Features



  • The system is fully food grade

  • No endotoxins are produced

  • No inclusion bodies

  • No spores

  • No extracellular proteases

  • Tightly controlled gene expression allows production of toxic proteins

  • Simple fermentation, scale-up and downstream processing



Description



  • Next to the wealth of traditional food applications, L. lactis is
     increasingly used for modern biotechnological applications such as the
     production of recombinant proteins for food, feed, pharmacological and
     biocatalysis applications. Owing to the easy genetic accessibility of L.
     lactis
    the Nisin Controlled gene
    Expression (NICE®) system developed at NIZO has
     played a key role in this development.



Applications



  • Production of homologous and heterologous proteins for food, feed,
     pharmacological and biocatalysis applications

  • Production of prokaryotic and eukaryotic membrane proteins

  • Production of exopolysaccharides

  • Production of ingredients through metabolic engineering: e.g. alanine,
     folate, diacetyl

  • Preparation of L. lactis as a biocatalyst by expression of a suitable
     enzyme as, e.g., dehydrogenases and in-situ cofactor regeneration

  • High-throughput screening for enzyme evolution or enzyme comparison.




Plasmids for Intracellular Expression and Secretion

胞内表达和分泌表达乳酸菌载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心






Vector map of pNZ9530



Plasmids for Intracellular Expression



  • The replicons of the vectors pNZ8008, pNZ8148, pNZ8149 and pNZ8150 are
     identical and came originally from the Lactococcus lactis plasmid pSH71.
     However, this replicon has a broad host range. Plasmids with this replicon can
     replicate in many Gram-positive bacteria, such as Lactobacillus plantarum
     and Streptococcus thermophilus.



  • pNZ8148 – In this vector the nisA promoter is followed by an NcoI
     site for translational fusions at the ATG. It contains a terminator after the
     MCS. Sequence adaptation for cloning in NcoI can result in a change of the
     second amino acid of a protein (Mierau and Kleerebezem, 2005).



  • pNZ8149 – This vector has the lacF gene as food grade
     selection marker. To enable selection of transformants, it needs a host strain
     with the lactose operon without the lacF gene like Lactococcus
     lactis
    NZ3900. The nisA promoter is followed by an NcoI site for
     translational fusions at the ATG. It contains a terminator after the MCS.
     Sequence adaptation for cloning in NcoI can result in a change of the second
     amino acid of a protein (no reference, example: (Mierau et al., 2005)).



  • pNZ8150 – The nisA promoter is followed by a ScaI site for
     translational fusions precisely at the ATG. Blunt end fragments are generated by
     PCR and cannot be cut out again after ligation to the ScaI site (Mierau and
     Kleerebezem, 2005).



  • pNZ8151 – This vector carries the lacF gene as food grade
     selection marker. The nisA promoter is followed by an ScaI site for
     translational fusions at the ATG start codon. It contains a terminator after the
     MCS (Mierau and Kleerebezem, 2005).



  • pNZ8152 – This vector contains the alr gene as food grade
     selection marker. The nisA promoter is followed by an ScaI site for
     translational fusion at the ATG start codon. It contains a terminator after the
     MCS (Mierau and Kleerebezem, 2005).



  • pNZ8008 – This is a reference plasmid for testing the nisin induction
     in Lactococcus and other lactic acid bacteria genera. A gusA gene
     without promoter was fused to the nisin A promoter (PnisA) (de Ruyter et
     al.
    , 1996).



  • pNZ9530 – A low copy plasmid with pAMΒ1 origin of replication that
     carries the nisR and nisK genes. For cloning in Lactococcus
     strains and in strains of other lactic acid bacteria genera that do not have the
     regulatory genes integrated into the chromosome. In this case, a two plasmid
     system is used for nisin induced expression: e.g. pNZ9530 (nisRnisK) + pNZ8150
     (+insert) (Kleerebezem et al., 1997).



Secretion Vectors



  • The secretion vectors pNZ8120, pNZ8121, pNZ8122, pNZ8123, and pNZ8124 are
     broad host range vectors with a chloramphenicol selection marker. They differ in
     their signal sequence and in their cloning sites.



  • pNZ8120 – A vector with the signal sequence of the lactococcal cell
     wall proteinase PrtP (Vos et al.). Cloning is possible via an NaeI site
     and the mature protein starts with its own first amino acid (results not
     published).



  • pNZ8121 – This vector has the signal sequence of PrtP also, but
     cloning is done via an EcoRV site, so that the mature protein starts with the
     first amino acid of PrtP after the signal cleavage site. This setting can
     contribute to a greater efficiency in removing the signal peptide (results not
     published).



  • pNZ8122 – A vector with the signal sequence of the Lactobacillus
     brevis
    SlpA protein (Novotny et al., 2005). Cloning is done via an
     NruI site, so the mature protein starts with the first amino acid of SlpA after
     the signal cleavage site. This setting can contribute to a greater efficiency in
     removing the signal peptide (results not published).



  • pNZ8123 – This vector provides the signal sequence of the lactococcal
     major secreted protein Usp45. Cloning is done via an NaeI site; mature protein
     starts with its own first amino acid (results not published).



  • pNZ8124 – Like pNZ8123 this vector has the signal sequence of Usp45,
     but cloning is done via an EcoRV site and the mature protein starts with the
     first amino acid of Usp45 (van Asseldong et al., 1993 & 1990) after
     the signal cleavage site. This setting can contribute to a greater efficiency in
     removing the signal peptide (results not published).



Overview on available Plasmids










































































































Host Strains




Electron microscope image of Lactococcus lactis



Description



  • Lactococcus lactis NZ9000pepN::nisRnisK;

  • Standard host strain for nisin regulated gene expression (NICE®).
     The strain contains the regulatory genes nisR and nisK integrated
     into the pepN gene (broad range amino peptidase) (Kuipers et al.,
     1998; Mierau and Kleerebezem, 2005).



  • Lactococcus lactis NZ9100nisRnisK;

  • Standard host strain for nisin regulated gene expression (NICE®).
     The strain contains the regulatory genes nisR and nisK integrated
     in a neutral locus.



  • Lactoccoccus lactis NZ9130alr-, nisRnisK; This host
     strain is the same as N9100 used for nisin regulated gene expression. It carries
     an alr deletion (Δalr) that encodes for alanine racemase. Deletion
     of the alr gene results in auxotrophy for alanine suggesting that the
     strain is unable to grow on media without alanine unless alr is provided
     on a plasmid (Bron et al., 2002).



  • Lactococcus lactis NZ3900lacF-, pepN::nisRnisK;
     Standard strain for food grade selection based upon the ability to grow on
     lactose. This strain is a progeny of NZ3000, a strain in which the lactose
     operon, that is generally present on plasmids, has been integrated into the
     chromosome and the lacF gene was deleted. Deletion of the lacF gene makes
     this strain unable to grow on lactose unless lacF is provided on a plasmid (de
     Ruyter et al., 1996a).



  • Lactococcus lactis NZ3910lacF-, nisRnisK;

  • Same as NZ3900 for food grade lacF selection, but with
    nisRnisK integrated in a neutral locus.



Overview on L. lactis Host Strains







































StrainOrder #SpeciesFood GradeRemarks
NZ9000NTCCS09000-01L. lactisNOpepN::nisRnisK
NZ9100NTCCS09100L. lactisNOnisRnisK in neutral locus
NZ9130NTCCS09130-01L. lactisYESnisRnisK in neutral locus, alr-
NZ3900NTCCS03900-01L. lactisYESpepN::nisRnisK, lacF-
NZ3910NTCCS03910-01L. lactisYESnisRnisK in neutral locus, lacF-

您正在向 biovector.net  发送关于产品 Nisin诱导调控乳酸菌表达载体系统及宿主菌-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心 的询问

点击“立即发送”后,我们将在1个工作日内与您取得联系。