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引用和文献 (9)
Invitrogen™
BaculoDirect™ C 端表达试剂盒
The BaculoDirect™ Baculovirus Expression System is a powerful and versatile eukaryotic system for high-level protein expression in insect cells. The了解更多信息
| 货号 | 数量 |
|---|---|
| 12562013 | 5 transfections |
货号 12562013
价格（CNY)
-
数量:
5 transfections
The BaculoDirect™ Baculovirus Expression System is a powerful and versatile eukaryotic system for high-level protein expression in insect cells. The combination of Gateway™ Technology with baculovirus expression makes the BaculoDirect System the fastest and easiest method for production of recombinant baculovirus.
How it works
BaculoDirect™ Linear DNA (the baculovirus genome) is engineered to include attR sites for quick and efficient recombination with a Gateway entry clone. The gene of interest is recombined from the entry clone into the BaculoDirect Linear DNA using a simple, one-hour LR reaction (Figure 1). The resulting reaction mix contains the recombinant baculovirus carrying the gene of interest and is used to transfect insect cells. The need for transforming bacteria and isolating a large bacmid or co-transfecting a transfer vector and linear baculovirus DNA into insect cells is eliminated. As a result, the hands-on time is greatly reduced. Purified baculovirus can be isolated in less than one week.
Gateway linear DNA
BaculoDirect Linear DNA is designed for rapid cloning with a Gateway entry clone and subsequent expression in Sf9 or Sf21 insect cells. The Linear DNA features:
• Strong polyhedrin promoter for high-level expression
• R-sites for efficient recombination with any attL-flanked Gateway entry vector
• TK gene for negative selection using ganciclovir
• C-terminal V5-His tag (BaculoDirect™ C-Term Expression Kit) or N-terminal V5-His tag (BaculoDirect™ N-Term Expression Kit) for detection with anti-V5 antibody and purification with nickel-chelating resin
• TEV protease cleavage site for removal of the V5-His tag following purification (BaculoDirect N-Term Expression Kit)
• LacZ gene, to ensure a pure baculovirus stock is generated, which is replaced by your gene of interest after LR recombinant reaction
Additional materials required, available separately: Gateway entry clone.
A selection guide for choosing the most appropriate Gateway entry vector for your application can be found at: www.thermofisher.cn/Gateway.
How it works
BaculoDirect™ Linear DNA (the baculovirus genome) is engineered to include attR sites for quick and efficient recombination with a Gateway entry clone. The gene of interest is recombined from the entry clone into the BaculoDirect Linear DNA using a simple, one-hour LR reaction (Figure 1). The resulting reaction mix contains the recombinant baculovirus carrying the gene of interest and is used to transfect insect cells. The need for transforming bacteria and isolating a large bacmid or co-transfecting a transfer vector and linear baculovirus DNA into insect cells is eliminated. As a result, the hands-on time is greatly reduced. Purified baculovirus can be isolated in less than one week.
Gateway linear DNA
BaculoDirect Linear DNA is designed for rapid cloning with a Gateway entry clone and subsequent expression in Sf9 or Sf21 insect cells. The Linear DNA features:
• Strong polyhedrin promoter for high-level expression
• R-sites for efficient recombination with any attL-flanked Gateway entry vector
• TK gene for negative selection using ganciclovir
• C-terminal V5-His tag (BaculoDirect™ C-Term Expression Kit) or N-terminal V5-His tag (BaculoDirect™ N-Term Expression Kit) for detection with anti-V5 antibody and purification with nickel-chelating resin
• TEV protease cleavage site for removal of the V5-His tag following purification (BaculoDirect N-Term Expression Kit)
• LacZ gene, to ensure a pure baculovirus stock is generated, which is replaced by your gene of interest after LR recombinant reaction
Additional materials required, available separately: Gateway entry clone.
A selection guide for choosing the most appropriate Gateway entry vector for your application can be found at: www.thermofisher.cn/Gateway.
仅供科研使用。不可用于诊断程序。
规格
产品类型表达试剂盒
数量5 transfections
载体BaculoDirect 载体
克隆方法Gateway™
产品线BaculoDirect、Gateway, Gateway
促进剂多角体蛋白
蛋白标记His 标签 (6x)、V5 抗原决定簇标签, V5 Epitope Tag
Unit SizeEach
内容与储存
BaculoDirect 表达试剂盒包括 5 个小瓶（0.3 µg/瓶）的 BaculoDirect 线性 DNA、125 µL Cellfectin 转染试剂、冷冻 Sf9 细胞、Gateway LR Clonase 酶混合物、GIBCO 无添加剂的 Grace 昆虫培养基和更昔洛韦。
在 +4°C 下储存 BaculoDirect 线性 DNA、Cellfectin 试剂和 Grace 培养基。在 -80°C 下储存 LR Clonase 混合物。将 Sf9 细胞储存于液氮中。如果妥善储存，可保证所有组分稳定储存 6 个月。
在 +4°C 下储存 BaculoDirect 线性 DNA、Cellfectin 试剂和 Grace 培养基。在 -80°C 下储存 LR Clonase 混合物。将 Sf9 细胞储存于液氮中。如果妥善储存，可保证所有组分稳定储存 6 个月。
常见问题解答 (FAQ)
白色菌落不能在液体培养基中生长。我该怎么办？
为什么会出现蓝色菌落？为什么会出现中心为蓝色而边缘为白色的菌落？
我得到的主要是白色/野生型空斑，而不是蓝色/重组型空斑。哪里出错了？
感染细胞后，我在一个细胞中看到较大的多角体，而在相邻细胞中看到较小的多角体（数量更多）。这正常吗？
我担心得不到空斑。出现空斑需要几天？空斑通常为多大？
引用和文献 (9)
引用和文献
Abstract
Functional characterization of five novel CYP2C8 variants, G171S, R186X, R186G,
K247R, and K383N, found in a Japanese population.
Journal:Drug Metab Dispos
PubMed ID:15716363
Cytochrome P450 2C8 is one of the primary enzymes responsible for the metabolism
of a wide range of drugs such as paclitaxel, cerivastatin, and amiodarone. We have sequenced the CYP2C8
gene from 201 Japanese subjects and found five novel nonsynonymous single nucleotide polymorphisms (SNPs):
511G>A (G171S), 556C>T (R186X; X represents the translational stop
Analysis of the large inactive P-TEFb complex indicates that it contains one 7SK molecule, a dimer of HEXIM1 or HEXIM2, and two P-TEFb molecules containing Cdk9 phosphorylated at threonine 186.
Journal:J Biol Chem
PubMed ID:15965233
'Positive transcription elongation factor b (P-TEFb) regulates eukaryotic gene expression at the level of elongation, and is itself controlled by the reversible association of 7SK RNA and an RNA binding protein, HEXIM1 or HEXIM2. To further understand how P-TEFb is regulated, we analyzed the stoichiometry of all the known components
HEXIM2, a HEXIM1-related protein, regulates positive transcription elongation factor b through association with 7SK.
Journal:J Biol Chem
PubMed ID:15713662
'The kinase activity of positive transcription elongation factor b (P-TEFb), composed of cyclin-dependent kinase 9 and cyclin T1 or T2, is required for the transition of RNA polymerase II into productive elongation. P-TEFb activity has been shown to be negatively regulated by association with the small nuclear RNA 7SK and
Replication-dependent destruction of Cdt1 limits DNA replication to a single round per cell cycle in Xenopus egg extracts.
Journal:Genes Dev
PubMed ID:15598982
'In eukaryotes, prereplication complexes (pre-RCs) containing ORC, Cdc6, Cdt1, and MCM2-7 are assembled on chromatin in the G1 phase. In S phase, when DNA replication initiates, pre-RCs are disassembled, and new pre-RC assembly is restricted until the following G1 period. As a result, DNA replication is limited to a single
Characterization of Aplysia enticin and temptin, two novel water-borne protein pheromones that act in concert with attractin to stimulate mate attraction.
Journal:J Biol Chem
PubMed ID:15054104
'Mate attraction in Aplysia involves a long-distance water-borne signal (attractin) that is released during egg laying. Other pheromones are predicted to be released during egg laying that act in concert with albumen gland attractin to stimulate attraction, but their identities are unknown. To identify other candidate water-borne pheromones, we employed