产品简介：
TP53基因敲除质粒（pLenti-TP53-sgRNA ）以U6为启动子，可以在动物细胞中同时表达Cas9、目的基因的sgRNA和puromycin抗性基因。用于在动物细胞中直接基于CRISPR/Cas9技术敲除目的基因，或者与pMDLg、Rev及VSV-g共转HEK293T细胞，通过包装慢病毒后基于CRISPR/Cas9技术敲除目的基因。本质粒全长约13,000bp，关键图谱信息请参考下图。
本质粒为氨苄青霉素抗性和嘌呤霉素(Puromycin)抗性，转化大肠杆菌后，可使用氨苄青霉素抗性筛选阳性菌；转染细胞后，可以使用嘌呤霉素筛选稳定表达融合蛋白的细胞株。
TP53基因的基本信息：具体全序列请详询技术支持
| Gene Symbol | TP53 |
| Official Full Name | tumor protein p53 |
| Species | Human |
| Gene ID | 7157 |
| GenBank Accession | BC003596 |
| Transcript | NM_000546 |
| Gene Summary | This gene encodes tumor protein p53, which responds to diverse cellular stresses to regulate target genesthat induce cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. p53 protein isexpressed at low level in normal cells and at a high level in a variety of transformed cell lines, where it’sbelieved to contribute to transformation and malignancy. p53 is a DNA-binding protein containingtranscription activation, DNA-binding, and oligomerization domains. It is postulated to bind to ap53-binding site and activate expression of downstream genes that inhibit growth and/or invasion, andthus function as a tumor suppressor. Mutants of p53 that frequently occur in a number of different humancancers fail to bind the consensus DNA binding site, and hence cause the loss of tumor suppressoractivity. Alterations of this gene occur not only as somatic mutations in human malignancies, but also asgermline mutations in some cancer-prone families with Li-Fraumeni syndrome. Multiple p53 variants dueto alternative promoters and multiple alternative splicing have been found. These variants encode distinctisoforms, which can regulate p53 transcriptional activity. |
使用说明：
- 首次使用时请先取少量本质粒转化大肠杆菌，进行质粒小量、中量或大量抽提后再用于后续用途。抽提获得的质粒可以通过酶切电泳或测序方法进行鉴定。
- 推荐鉴定使用的引物序列如下：
菌落PCR：PCR产物约为700bp
Forward primer: 5´- TATACGATACAAGGCTGTTAGAGAG-3´
Reverse primer: 5´- ACTGTGGGCGATGTGCGCTCTG-3´，
测序鉴定：
hU6，Forward primer: 5´- ATGGACTATCATATGCTTACCGTA-3´
比对序列:
ATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCGATTTCTTGGCTTTATATATCTTGTGGAAAGGACGAAACACCGNNNNNNNNNNNNNNNNNNNNGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTG。其中N为sgRNA序列。
保存条件：-20℃保存。
运输条件：湿冰运输。
注意事项：
- 如果需要sgRNA与质粒序列，请在收货后向技术支持索取。
- 本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
- 为了您的安全和健康，请穿实验服并戴一次性手套操作。