摘要
本研究首次从鲤(Cyprinus carpio)中得到2种形式的14 kD载脂蛋白(Apo-14kD)和4种形式的载脂蛋白E(ApoE),分别为Apo-14kDa-a和Apo-14kDa-b(同源性为79%)、ApoE-a1和ApoE-a2(同源性为91%)以及ApoE-b1和ApoE-b2(同源性为90%)。系统学分析表明,Apo-14kDa-a与多数硬骨鱼的Apo-14kDa同源性高于与Apo-14kDa-b的同源性,而Apo-14kDa-b与草鱼(Ctenopharyngodon idellus)的Apo-14kDa同源性较高,提示二者在鲤科鱼类中的分化时间较早;ApoE-a和ApoE-b的分化时间也应该处于鲤科鱼类分化的早期阶段,随后进一步分化出4个同源基因。半定量RT-PCR结果显示Apo-14kDa-a基因在肝、头肾中大量表达,肠中微弱表达,其他组织无表达,Apo-14kDa-b基因只在肝表达;ApoE-a1基因和ApoE-a2基因只在肠表达;而ApoE-b1基因和ApoE-b2基因均在脑、皮肤、鳃、肝、精巢、肠、心脏、肌肉、体肾、头肾中表达,但在脾和卵巢发生表达分化。进一步的qRT-PCR检测发现6个基因在不同胚胎发育时期的表达模式也有较大不同,且在不同时期的表达均存在阶段差异性。另外,qRT-PCR分析证实Apo-14kDa基因和ApoE基因均有可能在鲤胚胎发育过程中发挥着重要且显著不同的作用。
Apolipoproteins, the protein moieties of lipoproteins, participate in physical activities in different forms. A 14 kDa apolipoprotein （Apo-14 kDa） and apolipoprotein E （ApoE） are thought to be associated with embryonic development in animals. We analyzed the gene structure and phylogenetic evolution of these two types of apoli- poprotein in the common carp （Cyprinus carpio）. In addition, we documented the pattern of expression using RT-PCR to provide insight into their function during the embryonic development of carp. We obtained two types of Apo-14 kDa and four types of ApoE, including Apo-14kDa-a and Apo-14kDa-b （identity=79%）, ApoE-al and ApoE-a2 （identity=91%）, and ApoE-bl and ApoE-b2 （identity=90%）. Phylogenetic analysis revealed that Apo-14 kDa-a has a close relationship with that of most other teleosts and that Apo-14 kDa-b has the closest relationship with that of grass carp. These relationships likely reflect the early differentiation of the family Cyprinidae, al- though ApoE is characterized by four homologous genes. The expression of carp Apo-14 kDa-a was highest in the liver and head-kidney, was very low in the intestine, and undetectable in the remaining tissues. Conversely, we were only able to detect expression of Apo-14 kDa-b in the liver. We observed tissue specific differences in the expression ofApo-14 kDa-a and Apo-14 kDa-b. ApoE-al and ApoE-a2 were only detected in the intestine whereas ApoE-bl and ApoE-b2 were present in the brain, skin, gill, liver, spermary, intestine, heart, muscle, body-kidney, and head-kidney. ApoE-bl was also present in the spleen and ApoE-b2 in the ovary. Our observations suggest there is considerable variation in the expression and function of ApoE. Furthermore, qRToPCR analysis suggests that the expression of these six genes varies throughout embryonic development, both within and among genes. This is consistent with the molecular phylogeny of these genes, qRT-PCR analysis suggests that Apo-14 kDa and ApoE play an important role throughout different stages of embryonic development in common carp.
出处
《中国水产科学》
CAS
CSCD
北大核心
2012年第5期741-755,共15页
Journal of Fishery Sciences of China
基金
国家863计划项目(2009AA10Z105
2011AA100401)
农业部公益性行业科研专项项目(200903045)
中央级公益性科研院所基本科研业务费专项资金项目(2009B002)
