Phosphatase, Alkaline, Shrimp

磷酸酶，碱性，虾

Company: United States Biological

Catalog#: P4071-05

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Detection of piggyBac-mediated Transposition by Splinkerette PCR in Transgenic Lines of Strongyloides ratti

Author:

Hongguang Shao and James B. Lok,

Date:

2014-01-05

[Abstract] Splinkerette PCR (spPCR) is a newly developed and efficient method to ascertain and characterize genomic insertion sites of transgenes. The method described in this protocol was successfully applied to confirm piggyBac transposon-mediated integration of transgenes into chromosomes of the parasitic nematode Strongyloides ratti. This work is described in detail in Shao et al. (2012) and presented here in a simplified diagram (Figure 1). Using this method, chromosomal loci of integration were determined based on target site and 5’- and 3’ flanking sequences. Therefore, spPCR can be a useful method to confirm integrative transgenesis in functional genomic studies of parasitic nematodes. Potter and Luo (2010) contains a protocol for use of spPCR to detect and map piggyBac ... [摘要] Splinkerette PCR（spPCR）是一种新发展和有效的方法来确定和表征转基因的基因组插入位点。该方案中描述的方法被成功地应用于确认转座子介导的转基因整合到寄生线虫类线虫的染色体中的piggyBac 。该工作在Shao等人（2012）中详细描述并在简化图（图1）中呈现。使用该方法，基于靶位点和5'和3'侧翼序列确定整合的染色体基因座。因此，spPCR可以是确认在寄生线虫的功能基因组研究中的整合转基因的有用方法。 Potter和Luo（2010）包含使用spPCR检测和定位 piggyBac 转座子介导的染色体整合在果蝇中的方案，并且是我们的方法的来源类固醇。在该参考文献中描述的splinkerette和piggyBac 特异性寡核苷酸可以在没有修饰的情况下用于类花青苷。对于感兴趣的读者，可以在Viney和Lok（2007）中发现寄生线虫的生物学的一般综述，以及基于方法的文章。 stercoralis作为实验模型，具有转基因的信息，可以在Lok（2007）中找到。

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Phosphatase, Alkaline, Shrimp

磷酸酶，碱性，虾

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