蜜蜂精子介导GFP基因转移研究

江西农业大学动物科技学院郭冬生、天津农学院张巧利和泉州师范学院模式生物研究中心孙亮先三位动物分子生物学研究工作者将pGFP-2质粒线性化,利用精子介导,通过人工授精技术导入意大利蜜蜂处女王．然后将授精王介绍到无王群中繁殖后代,其结果显示：试验群产生了一群绿色荧光阳性蜜蜂,他们对此阳性蜂群后代分析发现,1～2日龄幼虫能坳4到荧光,提取蜜蜂DNA进行PCR扩增到预想的片断。     

转基因鸡的制备方法探讨

由于鸡本身的众多优点,转基因鸡的研究逐渐受到了研究者的重视,简要介绍了转基因鸡的发展史,对各种转基因技术方法进行了比较和分析,提出了每种方法的优缺点,尤其在精子介导法上作了较为详细的论述,并展望了转基因鸡的应用前景。     

精子介导转基因动物的制备

近年来 ,通过显微注射DNA至孵育的卵母细胞原核或外源基因转染后的胚胎干细胞进行转基因动物的生产已取得了令人瞩目的成就。在过去的 1 0年中 ,以精子作载体制备转基因哺乳动物或脊椎动物也取得了一些不同程度的进展。这些技术主要包括 :直接将外源DNA与精子共孵育至成熟 ;提取分离的精子DNA或进行预处理至精子发育成熟 ;以及在辅助受精前分离精子细胞等。此外 ,一些显微注射技术 ,如在输精管内进行体内直接转染雄性生殖细胞 ;将体内转染的雄性生殖细胞植入已分离的雄性生殖细胞 ,再显微注射至受体的睾丸 ,这些技术也逐渐成熟起来。研究表明 ,通过体内、体外转染外源DNA的显微操作技术只需将雄性受体与野生型雌性交配就可产生出转基因的后代个体 ,同时也避免了辅助受精和胚胎操作带来的机械损伤 ,因此具有一定的优势。本文综述了精子介导转基因 (SMGT)技术的发展历程、研究现状及前沿进展。     

精子介导的转基因技术

精子介导的转基因技术是近十几年发展起来的一种新技术。从各个种的实验证明,精子有瞬间吸收外源DNA的能力。这一过程由一系列因子起重要调节作用。一种特异的DNA结合蛋白介导了外源DNA与精子的结合,同时精浆中存在一种因子起抑制两者结合的拮抗作用。CD4分子与外源基因的内化有关。内化的DNA可能与精子核支架结构（nuclear scaffold)结合,并整合或重排。但仍需要大量实验数据进一步证明是否产生真正可遗传的转基因后代,及如何提高转基因效率,以使这一方法得到普遍的推广及应用。     

转基因动物技术的研究进展

转基因动物是其基因组内稳定整合了所导入的外源基因的动物。目前转基因实验动物体系的研究,主要集中在导入方法和提高整合和表达效率两个方面。本文对这两个方面的进展作一综述。 Abstract：Transgenic animals are those whose genome have foreign genes integrated. Nowadays, researches concentrated in the ways of gene transfer and of improving efficiency of integration and expression.The paper has con cluded these two aspects.     

家蚕转基因方法的初步研究

为建立家蚕转基因研究中切实可行的外源基因导入方法、分别用显微注射法、精子介导法、脂质体法和压力渗透法将含有绿色荧光蛋白(gfp)基因的转座子载体和辅助质粒转入到家蚕的受精卵中。在后代中检测到发绿色荧光的蚕茧,用PCR方法检测到后代个体染色体中含有gfp基因,并比较了上述几种方法的优缺点,为进一步进行转基因家蚕的研究奠定了基础。     

精子介导转基因动物的研究进展

精子是高度分化的细胞,它具有潜在的结合外源DNA并在受精过程中将其转入到卵内的能力。随着受精卵的发育,外源基因将随机整合到受体基因组中,部分基因能在成体中表达,部分基因不仅能表达还能遗传给后代。一旦精子能表达外源基因,那么将很容易得到大量的转基因后代。因此,精子能携带外源基因入卵和产量丰富这两大特性使精子作为载体制备转基因动物成为一个简便的途径。精子载体法制备转基因动物无需昂贵的实验设备,亦无需专门的技术。因此,提出后就备受关注。介绍精子介导的转基因动物的制作原理及具体方法的研究进展。     

精子载体法制备转基因动物的研究进展

精子载体法是一种低成本、简单快速制备转基因动物的方法.外源基因主要通过两种方式实现基因整合,一种是直接整合到精子的基因组中;另一种是通过精子携带进入卵细胞,然后整合到受精卵的基因组中.尽管现在已经有很多利用精子载体法获得了转基因动物的研究,但是基于精子在受精过程中,结合、内化、以及转运外源基因等方面能力的差异,转染效率仍有待提高.就利用精子载体法制备转基因动物过程中,精子载体法制备转基因动物的分子机制和方法方面进行系统的阐述和分析.     

中华蜜蜂精子介导egfp基因转移

中华蜜蜂Apis cerana cerana是一种真社会性昆虫,也是我国重要的经济昆虫。本实验目的是为了检测精子是否可以作为载体将外源egfp基因介导转入中华蜜蜂。首先将雄蜂精子与线性化的质粒DNA共浴,然后通过人工授精技术将精子导入处女王,再对实验蜂群后代进行分析。结果显示EGFP蛋白在一群实验组蜂的1～2日龄小幼虫中表达较强,能检测到0.01%～0.02%荧光阳性小幼虫个体;通过PCR和RT-PCR技术分析,证实转入的外源egfp基因获得表达。实验结果表明精子载体法能够用于中华蜜蜂外源基因的转移和表达。     

piggyBac转座子及其在转基因昆虫中的应用

piggyBac是一种从粉纹夜蛾Trichoplusiani.中分离到的、具有TTAA插入位点特异性的DNA转座子。piggyBac可在昆虫基因组中准确切离,转化频率较高,并且不受宿主因子的限制,是目前转基因昆虫研究中应用最广的转座子载体。近年来的研究发现,piggyBac类转座子广泛分布于昆虫和其他生物基因组中。文章从piggyBac的结构、转座特性、在转基因昆虫中的应用以及piggyBac类转座子的分布等几个方面综述了piggyBac的研究进展。     

蜜蜂嗅觉相关蛋白的研究进展

蜜蜂是一类营社会性生活的昆虫,蜂群中的蜂王、工蜂和雄蜂相互依存,各司其职,共同维持着群体严密有序的生活。其中,嗅觉系统在它们的生存和繁衍过程中起到重要的作用。昆虫对气味物质的感受过程是非常复杂的,需要有多种蛋白的参与。研究蜜蜂的化学感受机制可以帮助我们更深入地了解蜜蜂特有的行为及生物学特性。本文重点综述了与蜜蜂嗅觉相关的3种蛋白质的生化特性、分子结构、基因表达及其生理功能等方面的研究进展,以期为今后开展相关研究工作提供理论参考。     

蜜蜂以色列急性麻痹病毒(IAPV)研究进展

2006年冬季至2007年春季,美国发生了大范围的蜜蜂突然消失现象,类似的情况也发生在欧洲的部分地区,引发了严重的授粉危机。科学家对此展开了深入细致的研究并推测以色列急性麻痹病毒(Israeli acute paralysisvirus,IAPV)很可能是蜂群崩溃失调病(colony collapse disorder,CCD)的致病因素。随着研究的深入,对CCD的致病因素及其发生机制有了更深入的认识,并趋向于认为多方面的致病因素可能导致了CCD的发生。本文综述了目前国内外对IAPV的研究进展。     

Advances in the timing of spring cleaning by the honeybee Apis mellifera in Poland

1. The honeybee Apis mellifera is of huge worldwide economic importance in the pollination of crops for human consumption. In recent years, honeybee populations have declined under pressure from diseases and pests. Climate change is increasingly being viewed as an additional threat to honeybees and yet only limited research has been carried out in this area. 2. This paper reports the advance of the first cleansing flight (‘spring cleaning’) of the honeybee in Poznań, Poland, i.e. flights to excrete faeces, over a month in the period 1985–2009. The timing of this flight is advanced not only by higher late winter/spring temperatures but also by higher temperatures in the previous summer and autumn. 3. This earlier activity gives hope that the reported earlier flowering of many native and cultivated species will not cause a pollination synchrony crisis.     

The influence of some honeybee products as a diet substitute on the different stages of Coccinella undecimpunctata L. in Egypt

Several natural diets of honeybee products were tested for rearing Coccinella undecimpunctata L. under laboratory conditions. Two groups of tested adult and larval insect have been fed on the tested diets, the first group were fed on the tested honey bee products diets without adding aphids, while the second group fed on the same honeybee products with aphids added. The oviposition period in the first group was significantly shorter than control. The tested diets supplied with royal jelly + aphid and/or pollen grains + aphid caused the longest oviposition period. Diets of aphid alone and bee honey with aphid recorded the shortest oviposition period. Fecundity of ladybird was the highest in the tested diets with aphids than diets without aphid and/or aphid diet alone. The durations of larval and pupal stages were significantly different when compared with the diets with or without aphid and/or aphid diet alone. Further results indicated that the diets of royal jelly caused the shortest duration in larval and pupal stages.     

Effect of DNA concentration on transgenesis rates in mice and pigs

A retrospective analysis of transgenesis rates obtained in seven pronuclear microinjection programs was undertaken to determine if a relationship existed between the amount of DNA injected and transgenesis rates in the pig. Logistic regression analysis showed that as the concentration of DNA injected increased from 1 to 10ng/l, the number of transgenics when expressed as a proportion of the number liveborn (integration rate) increased from 4% to an average of 26%. A similar relationship was found when the number of molecules of DNA injected per picolitre was analysed. No evidence was obtained to suggest either parameter influenced integration rate in mice when the same constructs were injected. The number of transgenics liveborn when expressed as a proportion of ova injected (efficiency rate), increased as DNA concentration increased up to 7.5ng/l and then decreased at 10ng/l for both species suggesting that at this concentration DNA (or possible contaminants) may have influenced embryo survival. The relationship between efficiency and the number of molecules injected per picolitre was complex suggesting that the concentration at which DNA was injected was a better determinant of integration and efficiency rates. In conclusion, the present study suggests that transgenes need to be injected at concentrations of between 5 and 10ng/l to maximise integration and efficiency rates in pigs.     

Resources and transgenesis techniques for functional genomics in Xenopus

Recent developments in genomic resources and high‐throughput transgenesis techniques have allowed Xenopus to ‘metamorphose’ from a classic model for embryology to a leading‐edge experimental system for functional genomics. This process has incorporated the fast‐breeding diploid frog, Xenopus tropicalis, as a new model‐system for vertebrate genomics and genetics. Sequencing of the X. tropicalis genome is nearly complete, and its comparison with mammalian sequences offers a reliable guide for the genome‐wide prediction of cis‐regulatory elements. Unique cDNA sets have been generated for both X. tropicalis and X. laevis, which have facilitated non‐redundant, systematic gene expression screening and comprehensive gene expression analysis. A variety of transgenesis techniques are available for both X. laevis and X. tropicalis, and the appropriate procedure may be chosen depending on the purpose for which it is required. Effective use of these resources and techniques will help to reveal the overall picture of the complex wiring of gene regulatory networks that control vertebrate development.     

多巴胺神经通路及其对蜜蜂行为的影响

蜜蜂复杂的社会行为受到了研究者的广泛关注,了解蜜蜂的学习、记忆、导航、信息传递等行为的神经分子基础,可对探索人类自身的脑科学和复杂社会行为的分子基础提供比较研究信息。本文综述了多巴胺的作用机制及其在蜜蜂行为中的作用,详细介绍了蜜蜂脑部的多巴胺受体,总结了蜜蜂脑部多巴胺水平的影响因子等,最后对进一步研究多巴胺神经通路对蜜蜂行为的作用及机制的前景做一展望。     

蜜蜂采集及其信息传递的行为机制研究进展

蜜蜂的采集行为是蜜蜂众多社会行为中一种较复杂的行为,涉及信息评估、信息传递、学习记忆及能量代谢等不同的行为过程。研究蜜蜂采集及信息交流系统的分子机制,不仅利于蜜蜂的理论研究和蜂产业的发展,还为人类语言及信息交流系统的研究提供借鉴。本文从行为、感觉基础及分子机制等不同研究层面,综述了近年来对蜜蜂采集及信息传递行为的研究进展,并提出了研究设想,以期为深入研究蜜蜂的采集行为及其信息传递行为的分子机制提供参考。