Predicting the age of mosquitoes using transcriptional profiles
Predicting the age of mosquitoes using transcriptional profiles
复制标题
DOI：
10.1038/nprot.2007.396
复制
发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
O'Neill, Scott L.
中科院分区：
文献类型：
--
作者：
Cook, Peter E.;Hugo, Leon E.;O'Neill, Scott L.
使用转录谱预测蚊子年龄是一种新的解决方案，确定现场捕获的蚊子的年龄的长期问题。雌蚊的年龄对传播一系列人类病原体至关重要。转录年龄分级协议，我们在这里开发的埃及伊蚊，主要是作为一种研究工具。根据从已知年龄的蚊子收集的转录数据进行年龄预测。使用定量逆转录酶-PCR（RT-PCR）相对于参考基因定量八个候选基因转录物的丰度。使用典型冗余分析来分析归一化基因表达（GE）测量以获得蚊子年龄的多变量预测因子。第一冗余变量和已知年龄之间的关系被用作校准模型。对野生捕获的蚊子进行归一化GE测量，然后使用该校准模型预测年龄。将蚊子饲养到特定年龄以获得校准数据可能需要长达40天。转录丰度的分子分析和随后的年龄预测，对于100个个体应该需要类似的3-5天。
The use of transcriptional profiles for predicting mosquito age is a novel solution for the longstanding problem of determining the age of field-caught mosquitoes. Female mosquito age is of central importance to the transmission of a range of human pathogens. The transcriptional age-grading protocol we present here was developed in Aedes aegypti, principally as a research tool. Age predictions are made on the basis of transcriptional data collected from mosquitoes of known age. The abundance of eight candidate gene transcripts is quantified relative to a reference gene using quantitative reverse transcriptase-PCR (RT-PCR). Normalized gene expression (GE) measures are analyzed using canonical redundancy analysis to obtain a multivariate predictor of mosquito age. The relationship between the first redundancy variate and known age is used as the calibration model. Normalized GE measures are quantified for wild-caught mosquitoes, and ages are then predicted using this calibration model. Rearing of mosquitoes to specific ages for calibration data can take up to 40 d. Molecular analysis of transcript abundance, and subsequent age predictions, should take similar to 3-5 d for 100 individuals.