蜜蜂体内有9种王浆蛋白基因(major royal jelly protein,MRJPs1～9),其中MRJPs1～5在蜂王浆中含量较高,是蜂王浆生物学功能的基础。MRJPs6～9在王浆中没有或含量极少,且功能未知。为研究非王浆蛋白组分的MRJP9的生物学功能,本研究用RT-...蜜蜂体内有9种王浆蛋白基因(major royal jelly protein,MRJPs1～9),其中MRJPs1～5在蜂王浆中含量较高,是蜂王浆生物学功能的基础。MRJPs6～9在王浆中没有或含量极少,且功能未知。为研究非王浆蛋白组分的MRJP9的生物学功能,本研究用RT-PCR的方法对意大利蜜蜂Apis mellifera ligustica Spinola不同组织,不同部位,不同级型样本中mrjp9的转录水平进行检测和定量。结果发现mrjp9在蜜蜂的幼虫、蛹和成年蜜蜂的各组织部位均广泛转录表达,但其在幼虫、蛹和刚出房的成年蜜蜂体内表达水平较低,而在成年采集蜂体内表达水平则较高,其表达与蜜蜂的发育时期有关。通过对在成年蜜蜂体内各组织部位的表达水平进行检测的结果显示该基因主要在蜜蜂的头、胸和王浆腺等组织部位的表达较高,其他组织部位表达较少。此外,该基因也在雄蜂和蜂王体内广泛表达,不受蜜蜂性别和级型的影响。这些结果说明mrjp9是一与蜜蜂发育有关的基因,可能与蜜蜂的行为发育和分工调控有关。展开更多
Royal jelly （R J） is a well-known bioactive substance. It contains large amounts of major royal jelly proteins （MRJPs）, which express growth-factor-like activity in several animal and human cell lines. However, th...Royal jelly （R J） is a well-known bioactive substance. It contains large amounts of major royal jelly proteins （MRJPs）, which express growth-factor-like activity in several animal and human cell lines. However, the question on whether MRJPs possess growth-factor-like activity on all types of cell cultures remains. In order to determine whether MRJPs can be used as an alternative to fetal bovine serum （FBS） in different types of human cell culture, the prolif- eration of the complex serum with different ratios of MRJPs/FBS （M/F） was evaluated on five cell lines： 293T, HFL-I, 231, HCT116, and Changliver using MTT （3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2H-tetrazolium bromide） assay. The proliferation activity of the combination of the complex M/F serum with cytokines on the test cell lines was also measured. The results demonstrated that the complex serum with M/F 6/4 possessed the highest proliferation activity similar to or in excess of FBS. However, no activity of complex medium with M/F 6/4 was observed in 231 cells, indicating a selectivity of MRJPs on cell types. Compared with the complex medium with M/F 6/4, the complex medium with M/F 6/4 together with two cytokines, epidermal growth factor （EGF） and insulin-transferrin-selenium （ITS）, pro- moted proliferations of Changliver, 293T, HCT116, and H FL-I by 18.73%-56.19% （P〈0.01）. Our findings demonstrate that MRJPs could partially replace FBS in culturing many human cell lines.展开更多
蜂王浆主蛋白1(major royal jelly protein 1,MRJP1)是蜂王浆主蛋白家族中最重要的成员之一,MRJP1存在单体和低聚体两种形式。为了研究MRJP1低聚体的二级结构信息,首先利用ToyoScreen GigaCap Q-650M离子交换柱从新鲜蜂王浆中纯化出MRJP...蜂王浆主蛋白1(major royal jelly protein 1,MRJP1)是蜂王浆主蛋白家族中最重要的成员之一,MRJP1存在单体和低聚体两种形式。为了研究MRJP1低聚体的二级结构信息,首先利用ToyoScreen GigaCap Q-650M离子交换柱从新鲜蜂王浆中纯化出MRJP1低聚体,经分析超速离心和高效液相色谱-串联质谱技术鉴定其为MRJP1低聚体后,再通过圆二色谱技术检测MRJP1低聚体在不同浓度Ca2+条件下(10、50、200 mmol/L)二级结构的变化情况,并测定其变性温度。结果表明:该分离方法可以一步纯化出MRJP1低聚体,MRJP1低聚体的二级结构中β-折叠占比最高(55%左右),其次是无规卷曲(20%左右)和α-螺旋(15%左右),β-转角(10%左右)含量最低;MRJP1低聚体的变性温度为55℃。实验提出了MRJP1低聚体新的纯化方法及其二级结构的基本信息。展开更多
蜂王浆是决定蜜蜂幼虫发育中级型分化,即成为蜂王还是工蜂的关键环境因素,而蜂王浆主蛋白(main royal jelly proteins,MRJPs)是反映蜂王浆新鲜度的重要指标。日本镰仓昌树以蜜蜂和果蝇为模型的最新研究表明,MRJP1是蜂王浆中决定蜜蜂级...蜂王浆是决定蜜蜂幼虫发育中级型分化,即成为蜂王还是工蜂的关键环境因素,而蜂王浆主蛋白(main royal jelly proteins,MRJPs)是反映蜂王浆新鲜度的重要指标。日本镰仓昌树以蜜蜂和果蝇为模型的最新研究表明,MRJP1是蜂王浆中决定蜜蜂级型分化的关键因子,该蛋白可通过激活虫体脂肪体中的表皮生长因子信号通路,引发个体增大、发育时间缩短和卵巢发育等蜂王特征的出现。因此,今后很有必要进一步开展MRJP1对人体的营养功能和作用机理研究,为MRJP1应用于功能食品提供科学依据。展开更多
Major royal jelly protein 1（MRJP1）, designated apalbumin 1, has been regarded as a freshness marker of royal jelly（RJ）. A MRJP1-specific peptide（IKEALPHVPIFD） identified by bioinformatics analysis of homologous ...Major royal jelly protein 1（MRJP1）, designated apalbumin 1, has been regarded as a freshness marker of royal jelly（RJ）. A MRJP1-specific peptide（IKEALPHVPIFD） identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody（antiSP-MRJP1 antibody）. Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1（anti-R-MRJP1 antibody） reacted with other members of MRJP family in RJ. Enzyme-linked immunosorbent assay（ELISA） using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ（0 d）. Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage（P〈0.0001）. Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.展开更多
By screening the worker （Apis cerana cerana） heads cDNA library using a fragment of the mrjp3 gene of Apis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 （AccMRJP3）...By screening the worker （Apis cerana cerana） heads cDNA library using a fragment of the mrjp3 gene of Apis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 （AccMRJP3） cDNA was selected. Based on the sequencing of the inserts of the positive clone, a sequence of AccMRJP3 cDNA which is 1 887 bp long including a poly （A） tail was obtained. The AccMRJP3 cDNA encompassed an open-reading frame （ORF） with 1 779 bp encoding 593 amino acids. The un-translated regions （UTR） of the 5′ end and 3′end are 46 bp and 160 bp in length, respectively. Similar to AmMRJP3 and AdMRJP3, the putative AccMRJP3 also has a repetitive region. The comparison of the repetitive region of AccMRJP3, AmMRJP3 and AdMRJP3 shows some differences between them.展开更多
基金Project supported by the National Natural Science Foundation of China(No.31271848)the Important Scientific&Technical Innovation Project of Hangzhou(No.20131812A25)the Foundation of Fuli Institute of Food Science of Zhejiang University(No.KY201404),China
文摘Royal jelly （R J） is a well-known bioactive substance. It contains large amounts of major royal jelly proteins （MRJPs）, which express growth-factor-like activity in several animal and human cell lines. However, the question on whether MRJPs possess growth-factor-like activity on all types of cell cultures remains. In order to determine whether MRJPs can be used as an alternative to fetal bovine serum （FBS） in different types of human cell culture, the prolif- eration of the complex serum with different ratios of MRJPs/FBS （M/F） was evaluated on five cell lines： 293T, HFL-I, 231, HCT116, and Changliver using MTT （3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2H-tetrazolium bromide） assay. The proliferation activity of the combination of the complex M/F serum with cytokines on the test cell lines was also measured. The results demonstrated that the complex serum with M/F 6/4 possessed the highest proliferation activity similar to or in excess of FBS. However, no activity of complex medium with M/F 6/4 was observed in 231 cells, indicating a selectivity of MRJPs on cell types. Compared with the complex medium with M/F 6/4, the complex medium with M/F 6/4 together with two cytokines, epidermal growth factor （EGF） and insulin-transferrin-selenium （ITS）, pro- moted proliferations of Changliver, 293T, HCT116, and H FL-I by 18.73%-56.19% （P〈0.01）. Our findings demonstrate that MRJPs could partially replace FBS in culturing many human cell lines.
文摘蜂王浆主蛋白1(major royal jelly protein 1,MRJP1)是蜂王浆主蛋白家族中最重要的成员之一,MRJP1存在单体和低聚体两种形式。为了研究MRJP1低聚体的二级结构信息,首先利用ToyoScreen GigaCap Q-650M离子交换柱从新鲜蜂王浆中纯化出MRJP1低聚体,经分析超速离心和高效液相色谱-串联质谱技术鉴定其为MRJP1低聚体后,再通过圆二色谱技术检测MRJP1低聚体在不同浓度Ca2+条件下(10、50、200 mmol/L)二级结构的变化情况,并测定其变性温度。结果表明:该分离方法可以一步纯化出MRJP1低聚体,MRJP1低聚体的二级结构中β-折叠占比最高(55%左右),其次是无规卷曲(20%左右)和α-螺旋(15%左右),β-转角(10%左右)含量最低;MRJP1低聚体的变性温度为55℃。实验提出了MRJP1低聚体新的纯化方法及其二级结构的基本信息。
文摘蜂王浆是决定蜜蜂幼虫发育中级型分化,即成为蜂王还是工蜂的关键环境因素,而蜂王浆主蛋白(main royal jelly proteins,MRJPs)是反映蜂王浆新鲜度的重要指标。日本镰仓昌树以蜜蜂和果蝇为模型的最新研究表明,MRJP1是蜂王浆中决定蜜蜂级型分化的关键因子,该蛋白可通过激活虫体脂肪体中的表皮生长因子信号通路,引发个体增大、发育时间缩短和卵巢发育等蜂王特征的出现。因此,今后很有必要进一步开展MRJP1对人体的营养功能和作用机理研究,为MRJP1应用于功能食品提供科学依据。
基金supported by the Public Beneficial Scientific&Technical Plan of Zhejiang(No.2011C22039)the Important Scientific & Technical Plan of Zhejiang(No.2011C12023)+2 种基金the Important Scientific & Technical Innovation Project of Hangzhou(No.20131812A25)the Foundation of Fuli Institute of Food Science of Zhejiang University(No.KY201404)the National Natural Science Foundation of China(No.31271848)
文摘Major royal jelly protein 1（MRJP1）, designated apalbumin 1, has been regarded as a freshness marker of royal jelly（RJ）. A MRJP1-specific peptide（IKEALPHVPIFD） identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody（antiSP-MRJP1 antibody）. Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1（anti-R-MRJP1 antibody） reacted with other members of MRJP family in RJ. Enzyme-linked immunosorbent assay（ELISA） using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ（0 d）. Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage（P〈0.0001）. Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.
基金the National Natural Science Foundation of China(30200206) Zhejiang Provincial Natural Science Foundation of China(302113).
文摘By screening the worker （Apis cerana cerana） heads cDNA library using a fragment of the mrjp3 gene of Apis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 （AccMRJP3） cDNA was selected. Based on the sequencing of the inserts of the positive clone, a sequence of AccMRJP3 cDNA which is 1 887 bp long including a poly （A） tail was obtained. The AccMRJP3 cDNA encompassed an open-reading frame （ORF） with 1 779 bp encoding 593 amino acids. The un-translated regions （UTR） of the 5′ end and 3′end are 46 bp and 160 bp in length, respectively. Similar to AmMRJP3 and AdMRJP3, the putative AccMRJP3 also has a repetitive region. The comparison of the repetitive region of AccMRJP3, AmMRJP3 and AdMRJP3 shows some differences between them.
