正负选择系统 - 百度百科
胸苷激酶蛋白（tk）可使无毒的丙氧鸟苷（ganc，也称为鸟嘌呤）转变为毒性核苷酸，而杀死细胞，因而可用丙氧鸟苷筛选排除随机整合的细胞株。故同源重组时，g418和 ganc都有抗性，随机整合时对g418有抗性，但对ganc敏感，细胞将被杀死，无整合的将被g418杀死。
正负筛选（neo正向+HSV-tk负向）原理 - CSDN博客
2021年6月17日 · 在同源重组时，tk基因将被切除而丢失，相反在随机整合时，所有的序列均保留（包括tk）。胸苷激酶蛋白（TK）可使无毒的丙氧鸟苷（GANC，也称为鸟嘌呤）转变为毒性核苷酸，而杀死细胞，因而可用丙氧鸟苷筛选排除随机整合的细胞株。
[9] Use of fusions to thymidine kinase - ScienceDirect
2000年1月1日 · Full length HSV-tk can be used, complete from the first codon to the last. No amino acid linker (spacer) between HSV-tk and the second part of the protein seems to be necessary for the negative selection to function, at least in the cases tested so far.
正负筛选法 - 百度百科
构建一种特殊的 载体，该载体含有一段与靶基因同源的序列，在这段序列的某一外显子中插入Neor基因作为正选择标记；在同循序列之外的3’末端，或3’，5’两个末端接上 单纯疱疹病毒 的胸腺嘧啶激酶（HSV-tk）基因的序列作为负筛选标记。 经酶切后使其线性化，然后用电脉冲转染法导入细胞中，继续体外培养，并以药物G -418和GANC作双重筛选。 如果所导入的重组 DNA与受体细胞基因组DNA之间发生非同源重组，则外源基因通常是从头至尾均整合入受体细胞基因组中， …
Introduction of the Negative Selection Marker into Replacement …
1996年9月1日 · In this report, we present an alternative procedure to introduce any negative selection marker at the ends of a replacement vector using a simple ligation reaction, thereby avoiding laborious cloning procedures and plasmid size limitations.
New positive/negative selectable markers for mammalian cells on …
They are based on two genes for the resistance to Blasticidin S (BlaS) and on the thymidine kinase (Tk) gene of herpes simplex virus (HSV). The markers can be selected positively by their ability to induce BlaS resistance and negatively on the …
A new set of positive/negative selectable markers for …
1998年9月18日 · In the cases of Tk-BSD and Tk-bsr, the more effective the positive selection, the more GANC was required to kill all the cells. In conclusion, these five new markers function in eukaryotic cells and are selectable both positively and negatively.
The thymidine kinase/ganciclovir-mediated "suicide" effect is …
1995年6月12日 · The herpes simplex virus thymidine kinase (HSV-TK) converts ganciclovir (GCV) into a toxic product and allows selective elimination of TK+ cells in vitro and in vivo. It is currently being used in clinical gene therapy trials as a therapeutic gene or as a safety marker.
Multiplexing RMCE: Versatile Extensions of the Flp ... - ScienceDirect
2010年9月10日 · Ganc enables the survival only of cells that have lost the HSV-tk function—ideally those that underwent RMCE (cf. Fig. 7).
Structures of replacement vectors for efficient gene targeting
When structures of the vector DNA fragments present in 20 G418-resistant and GANC-resistant non-targeted clones were examined by Southern blot analysis, the inefficiency of GANC selection proved to be mostly caused by exonucleolytic degradation of HSV-tk genes progressing from ends of the vectors.