pAAV-U6-GFP腺相关病毒荧光表达载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pAAV-U6-GFP BioVector NTCC质粒载体菌种细胞基因保藏中心

pAAV-U6-GFP腺相关病毒荧光表达载体，带有mU6启动子，GFP荧光报告基因。Amp抗性。

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
电话：+86-010-53513060
网址：www.biovector.net
Adeno-associated viruses (AAVs) are derived from defective parvoviruses, which depend on essential
helper functions provided by other viruses, such as adenovirus and herpes virus, for efficient viral
replication and propagation. AAV has no etiologic association with any known diseases and has been
successfully used to establish efficient and long-term gene expression in vivo in a variety of tissues
without significant cellular immune responses or toxicity.
AAV has a single-stranded DNA genome which consists of approximately 4.7 kb. All characterized
AAV serotypesshare three key features, including two copies of AAV terminal repeats (ITRs), one rep
region and one cap region. The ITRs are capable of forming T-shape secondary structure and are the
only cis elements that are required for AAV replication, packaging, integration, and rescue. The rep
region encodes four overlapping proteins designated as Rep78, Rep68, Rep52, and Rep40, according to
the apparent molecular mass of the protein. In addition to their well-defined roles in AAV replication,
Rep proteins also regulate AAV packaging and site-specific integration. The cap region encodes three
structural proteins, VP1, VP2, and VP3. These three proteins share the same reading frame (see Figure
1).
MCS:
• Enzyme Sites: 5’- BamHI, EcoRI, SalI -3’
• MCS Sequence: AGCCGGATCCAACGAATTCAACGTCGACTTTT
Designing Oligonucleotides:
It is necessary to synthesize two complementary oligonucleotides (a top strand and a bottom strand) for
each shRNA target site (Figure 2). The key features of the oligonucleotides include:
• A 5'-BamH I restriction site overhang on the top strand and a 5'-EcoR I restriction site
overhang on the bottom strand. These restriction sites will enable directional cloning of the
annealed oligonucleotides into the pAAV U6-GFP vector.
• A guanine (G) residue added upstream of the 5'-end of the shRNA sense strand as Pol III
transcription start site, if the target sequence does not start with a purine.
• A 19-base oligonucleotide sense sequence (target sense sequence) of the shRNA target site.
• A 9-nucleotide hairpin loop sequence (5'-TTCAAGAGA- 3').
• The 19-base oligonucleotide antisense sequence (target antisense sequence) of the shRNA
target site.
• A RNA Pol III terminator sequence consisting of a 6-nucleotide poly(T) tract.
BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
电话：+86-010-53513060
网址：www.biovector.net      [Supplier来源]  http://www.biovector.net