家蚕末龄幼虫、蛹和成虫期载脂蛋白组成的变化(英文)

【目的】载脂蛋白是一种重要的血淋巴脂蛋白,它将90%的脂类输送到内部器官。本研究旨在调查家蚕Bombyx mori各个发育期血淋巴载脂蛋白组成变化。【方法】采用溴化钾密度梯度超离心、肝素钠沉淀(heparin sodium precipitation)和连接蒸发光散射检测器的高效液相色谱(high performance liquid chromatography coupled with evaporative light scattering detector,HPLC-ELSD)等方法,检测了家蚕各个发育期血淋巴脂蛋白组成变化。【结果】从5龄幼虫到成虫的转变过程中,从幼虫期和吐丝期家蚕中分离的载脂蛋白密度分别为1.130和1.139 g/mL,但蛹期载脂蛋白密度降低(为1.083 g/mL)。应用肝素钠沉淀法对载脂蛋白组成的分析表明,家蚕吐丝期载脂蛋白主要是高密度载脂蛋白(9.8μg/μL);从吐丝期开始,低密度载脂蛋白含量增加,并在蛹期持续存在(4.2μg/μL)。在所有发育期,载脂蛋白Ⅲ含量低,在2日龄蛹中含量最高(4.0μg/μL)。另外,从吐丝期至成虫期,总脂浓度逐渐增加,从1.4μg/μL增加至3.8μg/μL。通过HPLC-ELSD对中性脂组成的分析表明,在所有发育期只含有微量的单酰甘油(<0.04μg/μL),在5龄幼虫中期至蛹期末观察到二酰甘油(0.11~0.61μg/μL),在蛹期发现三酰甘油(0.18~1.02μg/μL)。【结论】总体结果说明,在幼虫期和吐丝期,高密度载脂蛋白主要运输二酰甘油;化蛹后,高密度载脂蛋白更多地与三酰甘油有关,而与二酰甘油相关性小,并与载脂蛋白Ⅲ形成低密度载脂蛋白有关。这些结果提示载脂蛋白的转变是一个精确的时间控制过程。

A simple and cost-effective protocol for extraction of genomic DNA from ethanol preserved black flies(Simuliidae:Diptera)

Objective:To investigate the efficacy of extraction methodology for the proposed DNA from the ethanol preserved black flies(Simuliidae:Diptera).Methods:This study addressed a simple and effective protocol for extraction of DNA from black flies stored in the ethanol.The sizes of larval and adult black flies ranged from 1.5 to 2.5 mm and 3 to 7 mm,respectively.To demonstrate the efficacy of the proposed methodology,the DNA was extracted from the ethanol preserved sample of black flies using the commercial kids.The extracted DNA was further validated in the PCR amplification using internal transcribed spacer-1 rDNA and cytochrome oxidase subunit II rDNA target primers.Results:Interestingly,a minor modification in the proposed methodology yielded a good DNA concentration in comparison with the commercial kids.The extracted DNA sample using the proposed methodology was successfully validated in the PCR amplification using internal transcribed spacer-1 rDNA and cytochrome oxidase subunit II rDNA genes.Conclusions:The proposed DNA extraction procedure yielded good concentration of DNA from the ethanol preserved black flies.The added advantage is that the procedure is suitable for a range of insect species preserved in the ethanol obtained from the various field conditions.