BG-11 Media, optimized for cyanobacteria, 1 L - FAQs

View additional product information for BG-11 Media, optimized for cyanobacteria - FAQs (A1379902, A1379901)

13 product FAQs found

使用pSyn_1载体时，目的基因的基础表达很低。你们有何建议？

pSyn_1的启动子是Psc。Psc是一种弱组成型启动子，可启动目的基因的基础表达。而pSyn_6的启动子是psbA启动子（PpsbA）。PpsbA是细长聚球藻psbA基因（编码光合系统II蛋白D1）的一种强组成型启动子，可启动目的基因的高水平表达。

pSyn_1与pSyn_6有何区别？

pSyn_1的启动子是Psc。Psc是一种弱组成型启动子，可启动目的基因的本底表达。该启动子来自集胞藻PCC6803的茄呢酰二磷酸酯合酶基因。注意：该弱组成型启动子适用于受强表达阻碍的应用，如通路基因工程或低水平表达型突变基因的互补作用（Simkovsky et al., 2012）。

pSyn_6的启动子是psbA启动子（PpsbA）。PpsbA是细长聚球藻psbA基因（编码光合系统II蛋白D1）的一种强组成型启动子，可启动目的基因的高水平表达。pSyn-6载体还包括一个用于有效起始翻译的核糖体结合位点（GAAGGAG）和一个终止密码子。载体骨架还包含一个6xHis TEV和一个V5-His附加表位，用于检测/纯化目的基因。两种载体都含有NS1（中性位点1）同源重组位点，用于将载体整合到细长聚球藻基因组中。

聚球藻细胞的保质期/有效期是多久？

聚球藻在保存6个月后不会丧失活力。我们尚未对超过6个月的保存时间进行测试。

使用聚球藻表达试剂盒进行整合的原理是什么？

在基因组中性位点1（Neutral Site 1）之间和质粒的NSa和NSb之间发生双重同源重组事件，从而将目的基因和壮观美素抗性整合到中性位点1内的基因组中。

你们的试剂盒中使用哪种聚球藻株？

使用的是Synechococcus elongatus PCC 7942，一种模型蓝藻。

Blue-Green-11（BG-11）培养基的配方是什么？

我们提供的专为蓝藻类优化的即用型培养基为BG-11（货号A1379901）。BG-11培养基有多种配方。请查看以下配方：

我们的BG-11培养基能够培养细长聚球藻（Synechococcus elongatus，PCC 7942株）。
项目 配方 (%w/v)
硼酸 H3BO3 0.00287%
四水氯化锰 MnCl2·4H2O 0.00181%
七水硫酸锌 ZnSO4·7H2O 0.00022%
二水钼酸钠 Na2MoO4 0.00039%
五水硫酸铜 CuSO4·5H2O 0.00008%
硝酸钠 NaNO3 0.15000%
二水氯化钙 CaCl2.·2H2O 0.00270%
枸橼酸铁铵（绿色） C6H5+4yFexNyO7 0.00120%
EDTA EDTA 0.00010%
磷酸氢二钾 K2HPO4 0.00390%
七水硫酸镁 MgSO4·7H2O 0.00750%
一水碳酸钠 Na2CO3·H2O 0.00200%

I'm getting low basal expression of my gene of interest when using the pSyn_1 vector. What do you suggest I try?

The promoter for pSyn_1 is the Psc. Psc is a weak constitutive promoter that drives basal expression of your gene of interest. The promoter for pSyn_6, on the other hand, is the psbA promoter (PpsbA). PpsbA is a strong constitutive promoter of the psbA gene (encoding photosystem II protein D1) from Synechococcus elongatus, driving the high level expression of your gene of interest.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What is the difference between pSyn_1 and pSyn_6?

The promoter for pSyn_1 is the Psc. Psc is a weak constitutive promoter that drives basal expression of your gene of interest. The promoter was taken from the solanesyl diphosphate synthase gene from Synechocystis sp. Strain PCC 6803. Note: Use of this weak constitutive promoter is a good fit for applications that are hindered by strong expression, such as pathway engineering or complementation of mutant genes normally expressed at low levels (Simkovsky et al., 2012).

The promoter for pSyn_6 is the psbA promoter (PpsbA). PpsbA is a strong constitutive promoter of the psbA gene (encoding photosystem II protein D1) from Synechococcus elongatus, driving high-level expression of your gene of interest. The pSyn-6 vector also includes a ribosome-binding site (GAAGGAG) for efficient initiation of translation, as well as a stop codon. A 6xHis TEV and a V5-His epitope tag is also included in the vector backbone for detection/purification of the gene of interest. NS1 (neutral site 1) homologous recombination sites are present in both vectors for the integration of the vector into the Synechococcus elongatus genome.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What is the shelf-life/competency of Synechococcus?

After 6 months of storage, the Synechococcus did not lose their competency. We have not yet tested longer storage points.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What is the mechanism of integration when using the Synechococcus expression kits?

A double homologous recombination event occurs between the Neutral Site 1 in the genome and between NSa and NSb in the plasmid. This results in the gene of interest and spectinomycin resistance integrating into the genome within the Neutral Site 1.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What is the strain of Synechococcus used in your kits?

We no longer offer any strains. Our reagents and protocols were developed using Synechococcus elongatus strain PCC 7942, a model cyanobacterium.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What is the formulation of Blue-Green-11 (BG-11) media?

We offer ready-to-use BG-11 Media, optimized for Cyanobacteria (Cat. No. A1379901). There are several BG11 media formulations. Please see our formulation below.

Our BG-11 media have been qualified for culturing Synechococcus elongatus (strain PCC 7942).
Item, Formula, (%w/v)
Boric Acid, H3BO3, 0.00287%
Manganese Chloride Tetrahydrate, MnCl2-4H2O, 0.00181%
Zinc Sulphate Heptahydrate, ZnSO4-7H2O, 0.00022%
Sodium Molybdate Dihydrate, Na2MoO4, 0.00039%
Copper Sulphate Pentahydrate, CuSO4-5H2O, 0.00008%
Sodium Nitrate, NaNO3, 0.15000%
Calcium Chloride Dihydrate, CaCl2-2H2O, 0.00270%
Ferric Ammonium Citrate (Green), C6H5 4yFexNyO7, 0.00120%
EDTA, EDTA, 0.00010%
Potassium Phosphate Dibasic, K2HPO4, 0.00390%
Magnesium Sulfate Heptahydrate, MgSO4-7H2O, 0.00750%
Sodium Carbonate Monohydrate, Na2CO3-1H2O, 0.00200%

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What does the GeneArt Synechococcus Protein Expression Vector include?

It includes the pSyn_6 vector. This vector has been tested for protein expression in the Synechococcus elongatus PPC 7942. You can get the Synechococcus elongatus PPC 7942 from academic resources (https://genome.jgi.doe.gov/portal/synel/synel.info.html) or from the ATCC Collection (https://www.atcc.org/products/all/33912.aspx).

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.