Abstract：
:Genetic cross is a powerful tool for studying malaria genes contributing to drug resistance, parasite development, and pathogenesis. Cloning and identification of recombinant progeny (RP) is laborious and expensive, especially when a large proportion of progeny derived from self-fertilization are present in the uncloned progeny of a genetic cross. Since the frequency of cross-fertilization affects the number of recombinant progeny in a genetic cross, it is important to optimize the procedure of a genetic cross to maximize the cross-fertilization. Here we investigated the factors that might influence the chances of obtaining RP from a genetic cross and showed that different Plasmodium yoelii strains/subspecies/clones had unique abilities in producing oocysts in a mosquito midgut. When a genetic cross is performed between two parents producing different numbers of functional gametocytes, the ratio of parental parasites must be adjusted to improve the chance of obtaining RP. An optimized parental ratio could be established based on oocyst counts from single infection of each parent before crossing experiments, which may reflect the efficiency of gametocyte production and/or fertilization. The timing of progeny cloning is also important; cloning of genetic cross progeny from mice directly infected with sporozoites (vs. frozen blood after needle passage) at a time when parasitemia is low (usually <1%) could improve the chance of obtaining RP. This study provides an optimized protocol for efficiently cloning RPs from a genetic cross of malaria parasites.
journal_name
Exp Parasitoljournal_title
Experimental parasitologyauthors
Qi Y,Zhu F,Li J,Fu Y,Pattaradilokrat S,Hong L,Liu S,Huang F,Xu W,Su XZdoi
10.1016/j.exppara.2012.10.011subject
Has Abstractpub_date
2013-01-01 00:00:00pages
44-50issue
1eissn
0014-4894issn
1090-2449pii
S0014-4894(12)00322-0journal_volume
133pub_type
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journal_title：Experimental parasitology
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journal_title：Experimental parasitology
pub_type： 杂志文章
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更新日期：2001-09-01 00:00:00
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pub_type： 杂志文章
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journal_title：Experimental parasitology
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pub_type： 杂志文章
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