Genome display and typing of Plasmodium parasites using anchored PolyA and PolyT oligonucleotides.
Genome display and typing of Plasmodium parasites using anchored PolyA and PolyT oligonucleotides.
复制标题
使用锚定 PolyA 和 PolyT 寡核苷酸对疟原虫寄生虫进行基因组展示和分型。
DOI：
10.1006/expr.2000.4492
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发表时间：
2000
影响因子：
2.1
通讯作者：
Carlton,JM
中科院分区：
文献类型：
--
作者：
Su,X;Carlton,JM
简单重复序列，如（A）n、（T）n或（TA）n是来自遗传杂交（Wellems et al. 1990）的独立后代的丰富DNA。当在恶性疟原虫富含AT的基因组中扩增时（Su和Wellems 1996）。存在简单序列的寡核苷酸，如polyT和polyA-使用引物（A）17 GTG，获得了几个PCR条带，其中两个可以通过连接到预期退火到基因组DNA中的多个位点而清楚地分配到染色体1和8，并且它们可以用于从遗传图谱的基因组微卫星标记扩增许多DNA片段（Su等人，1999）（图2）。这些结果表明，APAD可用于随机性遗传标记的鉴定.我们首先使用（T）20、（A）20或（TA）10的单一引物扩增恶性疟原虫基因组DNA。PCR产物的涂片方法类似于随机扩增多态性DNA（RAPD）（威廉姆斯（Williams）et.等人，1990; Postlethwait等人，1994）。在每种情况下获得（数据未显示）。为了降低PCR产物的复杂性，我们使用了锚定方法，其中还用来自A. p.的APAD引物扩增1至5个离散产物。
Simple sequence repeats, such as (A) n,(T) n, or (TA) n are abundant DNA from independent progeny of a genetic cross (Wellems et al. 1990) was examined for APAD segregation profiles. When amplified in the AT-rich genome of P. falciparum (Su and Wellems 1996). Oligonucleotides of simple sequences like polyT and polyA are there- with primer (A) 17GTG, several PCR bands were obtained, two of which could be clearly assigned to chromosomes 1 and 8 by linking to fore expected to anneal to multiple sites in the genomic DNA and they can be used to amplify numerous DNA fragments from the genome microsatellite markers of a genetic map (Su et al. 1999)(Fig. 2). These results showed that APAD can be used to identify genetic markers in an at random. We first amplified P. falciparum genomic DNA using a single primer of (T) 20,(A) 20, or (TA) 10. A smear of PCR products was approach similar to that described for Random Amplified Polymorphic DNA (RAPD)(Williams et. al. 1990; Postlethwait et al. 1994). obtained in each case (data not shown). To reduce the complexity of PCR products, we used an anchoring approach in which one to five Discrete products were also amplified with APAD primers from