三个意蜂种群MRJP3的基因表达及其蛋白含量的比较分析
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摘要
蜂王浆中的蛋白主要分为水溶性蛋白和水不溶性蛋白,其中水溶性蛋白占总蛋白含量的46%-89%,为王浆中的主要蛋白成分,即MRJPs(Major Royal Jelly Proteins)。研究表明,MRJPs为一组同源蛋白组成的蛋白系列,共有九种蛋白组成;且蜂王浆很多生物学活性功能,都与MRJPs密切相关。MRJP3作为MRJPs系列中的一种重要蛋白,近年来被证明具有多种生理功能,如参与抗过敏反应等,是王浆活性功能的重要体现之一。
本研究选取平湖浆蜂、苏王1号、美意三个产浆性能完全不同的种群为研究对象,首次利用荧光定量PCR技术比较mrjp3基因在三个种群的表达情况,并首次利用凝胶渗透色谱-激光光散射联用技术检测三种群工蜂生产蜂王浆中MRJP3的相对含量,并比较MRJP3在不同生产性能种群所产王浆中的差异,分析MRJP3的相对含量与蜂王浆产量之间的关系。同时利用SDS-PAGE电泳的方法分析当新鲜蜂王浆中MRJP3含量一致时,在一定温度下,MRJP3含量随贮存时间变化的规律。本研究主要结果如下:
1荧光定量PCR结果显示三种群间mrjp3基因实时表达无明显差异,存在基因表达的一致性。表明mrjp3基因表达在王浆生产性能明显不同的种群间没有差异,王浆高产基因表达对mrjp3基因没有影响。当蜂群群势一致时,蜂王浆中的MRJP3的绝对量可能保持一致。
2通过凝胶渗透色谱-激光光散射技术检测三种群蜂王浆中MRJP3相对含量结果显示三种群所产王浆MRJP3相对含量差异极显著,蜂王浆低产的美意种群MRJP3相对含量最高,高产的平湖浆蜂种群MRJP3相对含量最低,表明随着蜂王浆产量的不断上升,蜂王浆的品质呈现一定程度的降低。通过对蜂王浆产量与MRJP3相对含量的相关关系分析表明蜂王浆中MRJP3相对含量与蜂王浆生产性能回归公式:y=-0.8196x+231.41,单因素方差检验结果(P<0.05)表明MRJP3相对含量与其蜂王浆产量之间存在着显著的负相关。。
3首次利用SDS-PAGE电泳方法分析MRJP3在一定温度下随贮存时间的变化规律,结果表明,蜂王浆中主要蛋白主要由分子量在40kd-80kd之间的蛋白质组成,随贮存时间的延长,MRJP3发生不同程度的降解,相对含量不断降低。通过对蜂王浆贮存时间与MRJP3相对含量的相关分析表明蜂王浆中MRJP3相对含量与蜂王浆贮存时间回归公式y=-0.6446x+12.037,检验结果表明两者存着显著的负相关(P<0.05)。
Royal jelly proteins are divided into water insoluble proteins and water-soluble proteins ,which constitute about 46%-89% of the total protein content,identifying as major protein components of royal jelly ,that are MRJPs.It was found that MRJPs belong to one protein family consisting of a series of highly homologous proteins, which includes nine members and many functional biological activity of royal jelly are closely related with the it. in recent years, MRJP3 wihich as an important protein in the series of MRJPs has been shown to have many physiological functions ,such as participation in anti-allergenic ,is an important manifestation of rayal jelly in active funcion.
Pinghu pulp bee, American Italian bee and Suwang No.1 bee which have different performance in pulp production are selected in this study. The first time FQ-PCR to quantify the MRJP3 gene expression in the three populations, while using GPC - MALLS detection of three groups produced the relative concentration of royal jelly MRJP3 to explore the MRJP3 protein production performance in different populations which have different performance in pulp production, and whether as a measure of royal jelly freshness indicators to MRJP3.
At the same time SDS-PAGE protein electrophoresis are used to investigate royal jelly protein with normal temperature variation. The main results were as follows:
1. The same period, the same area ,the 10 days worker bee are selected in three different Italian honeybee population,the results of FQ-PCR show that :Three groups of relative gene expression of MRJP3 was no significant difference, indicating Italian bee production of royal jelly have no necessary link the level of performance to the gene expression of MRJP3 in individual worker bees.
2. GPC-MALLS be used to detect the MRJP3 protein concentration in the three strains,The results showed that: the MRJP3 protein concentration have significantly difference in the three strains. American Italian honeybee strains have the highest MRJP3 concentration, but pinghu pulp bee which have high-yield performance have the lowest MRJP3 concentrations.Contact FQ-PCR results,wo have the conclusion that MRJP3 gene for non-production genes, each worker bees have the same ablity to excrete the MRJP3,which not be used as indicators to measure the freshness of royal jelly.
3. SDS-PAGE protein electrophoresis be used to test royal jelly protein at room temperature with the variation of storage time,the results show that water-soluble royal jelly protein are composed of molecular weight of 40kd-80kd protein ,which gradually degraded into small molecules with the time.and the amount of protein degradation and storage time have the linear relation within a certain range.
本研究选取平湖浆蜂、苏王1号、美意三个产浆性能完全不同的种群为研究对象,首次利用荧光定量PCR技术比较mrjp3基因在三个种群的表达情况,并首次利用凝胶渗透色谱-激光光散射联用技术检测三种群工蜂生产蜂王浆中MRJP3的相对含量,并比较MRJP3在不同生产性能种群所产王浆中的差异,分析MRJP3的相对含量与蜂王浆产量之间的关系。同时利用SDS-PAGE电泳的方法分析当新鲜蜂王浆中MRJP3含量一致时,在一定温度下,MRJP3含量随贮存时间变化的规律。本研究主要结果如下:
1荧光定量PCR结果显示三种群间mrjp3基因实时表达无明显差异,存在基因表达的一致性。表明mrjp3基因表达在王浆生产性能明显不同的种群间没有差异,王浆高产基因表达对mrjp3基因没有影响。当蜂群群势一致时,蜂王浆中的MRJP3的绝对量可能保持一致。
2通过凝胶渗透色谱-激光光散射技术检测三种群蜂王浆中MRJP3相对含量结果显示三种群所产王浆MRJP3相对含量差异极显著,蜂王浆低产的美意种群MRJP3相对含量最高,高产的平湖浆蜂种群MRJP3相对含量最低,表明随着蜂王浆产量的不断上升,蜂王浆的品质呈现一定程度的降低。通过对蜂王浆产量与MRJP3相对含量的相关关系分析表明蜂王浆中MRJP3相对含量与蜂王浆生产性能回归公式:y=-0.8196x+231.41,单因素方差检验结果(P<0.05)表明MRJP3相对含量与其蜂王浆产量之间存在着显著的负相关。。
3首次利用SDS-PAGE电泳方法分析MRJP3在一定温度下随贮存时间的变化规律,结果表明,蜂王浆中主要蛋白主要由分子量在40kd-80kd之间的蛋白质组成,随贮存时间的延长,MRJP3发生不同程度的降解,相对含量不断降低。通过对蜂王浆贮存时间与MRJP3相对含量的相关分析表明蜂王浆中MRJP3相对含量与蜂王浆贮存时间回归公式y=-0.6446x+12.037,检验结果表明两者存着显著的负相关(P<0.05)。
Royal jelly proteins are divided into water insoluble proteins and water-soluble proteins ,which constitute about 46%-89% of the total protein content,identifying as major protein components of royal jelly ,that are MRJPs.It was found that MRJPs belong to one protein family consisting of a series of highly homologous proteins, which includes nine members and many functional biological activity of royal jelly are closely related with the it. in recent years, MRJP3 wihich as an important protein in the series of MRJPs has been shown to have many physiological functions ,such as participation in anti-allergenic ,is an important manifestation of rayal jelly in active funcion.
Pinghu pulp bee, American Italian bee and Suwang No.1 bee which have different performance in pulp production are selected in this study. The first time FQ-PCR to quantify the MRJP3 gene expression in the three populations, while using GPC - MALLS detection of three groups produced the relative concentration of royal jelly MRJP3 to explore the MRJP3 protein production performance in different populations which have different performance in pulp production, and whether as a measure of royal jelly freshness indicators to MRJP3.
At the same time SDS-PAGE protein electrophoresis are used to investigate royal jelly protein with normal temperature variation. The main results were as follows:
1. The same period, the same area ,the 10 days worker bee are selected in three different Italian honeybee population,the results of FQ-PCR show that :Three groups of relative gene expression of MRJP3 was no significant difference, indicating Italian bee production of royal jelly have no necessary link the level of performance to the gene expression of MRJP3 in individual worker bees.
2. GPC-MALLS be used to detect the MRJP3 protein concentration in the three strains,The results showed that: the MRJP3 protein concentration have significantly difference in the three strains. American Italian honeybee strains have the highest MRJP3 concentration, but pinghu pulp bee which have high-yield performance have the lowest MRJP3 concentrations.Contact FQ-PCR results,wo have the conclusion that MRJP3 gene for non-production genes, each worker bees have the same ablity to excrete the MRJP3,which not be used as indicators to measure the freshness of royal jelly.
3. SDS-PAGE protein electrophoresis be used to test royal jelly protein at room temperature with the variation of storage time,the results show that water-soluble royal jelly protein are composed of molecular weight of 40kd-80kd protein ,which gradually degraded into small molecules with the time.and the amount of protein degradation and storage time have the linear relation within a certain range.
引文
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