中国科学院昆明动物研究所机构知识库

果蝇是最经典的昆虫模式生物，摩尔根用果蝇发现了遗传学中的第三大定律——连锁互换定律，人们都说：“果蝇是上帝为摩尔根创造的”，但果蝇有何尝不是上帝送给科研工作者的礼物。从1933年到2011年，四次诺贝尔奖都出自果蝇，可以说是诺贝尔奖的摇篮。果蝇除了在经典遗传学方面做出的突出贡献以外，在在胚胎发育，器官形成，神经退化、肿瘤发生方面对人类也有很重要的借鉴意义，因为人类和果蝇的基因组中有80%的同源基因。除此之外果蝇的大脑也相当的复杂，除了学习、运动、交配等，还有选择、记忆、逃避等高层次行为能力，说明果蝇在研究神经生物学方面也会有很大的贡献，并且可以被人类借鉴。从摩尔根开始人们在遗传造作、制造突变个体等方面积累了大量的经验，已经把果蝇塑造成了一个非常成功且使用便利的模式生物，而蝴蝶中林奈把P.machaon作为模式种，但一直不能遗传操作，严重阻碍了蝴蝶这一极富进化意义类群的研究,我们在这里利用在果蝇中积累的经验，也对蝴蝶进行了遗传操作，着将是P.machaon成为蝴蝶中的模式生物的重要一步。最近几十年从头起源新基因在很多物种中被鉴定出来。而他们中有一些被认为是在基因组的改革和生物的演化方面都承担着重要的作用。在这个课题中我们敲除了黑腹果蝇中的两个从头起源新基因CG31955和CG32711，并且研究了寿命和生殖率等生物学功能。通过序列的分析和RNA以及蛋白水平的实验证据，证明这两个基因不仅真实存在，而且会表达翻译为蛋白，在黑腹果蝇生长发育的各个阶段行使功能。基因打靶造成突变后，这两个基因在在寿命和生殖方面都有明显的表型，而寿命和生殖是适应性进化的两个重要方面。CG31955被敲除之后发现雄性果蝇的生殖率降低了，而雌性果蝇的生殖力和寿命都有所增加。同时，CG32711被敲除后雌性雄性的生殖率都显著降低了，而雄性的寿命也变短了。这些数据显示，CG31955可能抑制了雌性黑腹果蝇的生殖率和寿命，但是会对雄性的生殖率有所俱进，CG32711对雌性和雄性的生殖率以及雄性的寿命有很重要的作用。通过对野生型和突变体的转录组分析，数据分析显示这两个从头起源新基因通过调控与几丁质和溶菌酶有关的基因，从而对雄性的生殖率有促进作用。CG31955对雌性可能是通过对产卵和溶菌酶有关的基因进行下，从而对生殖率和寿命进行抑制。CG32711可能是通过下调氧化磷酸化通路有关的基因去影响了雄性的寿命。我们的结果表明这两个从头起源新基因对生物体有着非常重要的作用。果蝇是很传统的模式生物，因为其易饲养，生长周期短而在生物研究中应用广泛，但是它可以解决的科学问题也比较有限。而蝴蝶因为它的多姿多彩一直让进化生物学家着迷，为什么蝴蝶可以进化出那么多的形态和色彩。随着近几年多种蝴蝶基因组的解析，基因编辑技术的发展，让蝴蝶成为了一个很有潜力的模式物种，尤其在研究演化、形态多样性的发展机制、以及物种分化方面。我们实验室前期解析的金凤蝶的基因组，被Linnaeus命名为所有蝴蝶的模式物种，很长时间以来人们一直在研究他的拟态、视觉体系、学习能力、和色素沉积的生物过程。CRISPR/Cas9基因编辑技术的出现，可以让人们在更广泛的方面去研究蝴蝶，就像其他模式生物：果蝇、家蚕一样，但还是不能确定这项技术是否能像在柑橘凤蝶中起作用一样，也在其他蝴蝶物种中作用。所以我们也就用相似的方法来在金凤蝶中进行基因编辑。我们的数据表明共同注射几个打靶位点的sgRNA进入金凤蝶的卵中，可以导致88.5%的幼虫形态发生变异。而且基因被破坏的比例达到了98.2%。实时定量PCR也表明在形态异常的个体中Abdominal-B基因表达量也变的非常低。这项工作表明，在柑橘凤蝶的形态发育过程也同样适用于其他蝴蝶物种，而且借助新的技术蝴蝶有巨大的潜力将会成新的研究遗传学、进化、以及发育的模型。家养动植物在人工选择下进化的遗传机制是一个古老而又悬而未决的生物学问题。1859年达尔文发表了其旷世著作《物种起源》。其第一章即题为“家养条件下的变异”，用人工选择的明显案例引证自然选择的伟大力量。山羊对人类来说算是驯化时间比较久的动物，在驯化的过程中很多的性状也发生了改变。比如,家山羊的毛质越来越好等等。我们利用荧光定量PCR随机检测了5个家山羊和野山羊基因拷贝数的差异。结果发现有四个基因在家山羊中的基因拷贝数比野山羊中的多。这样的结果为研究驯化过程中遗传物质的改变提供了更多的证据。 

Many de novo genes were recently identified from various species in the past decade. A few of them were investigated functionally, with evidence of performing important functions in genetic innovations and organismal evolution. Here, we investigated the biological roles and the underlying molecular mechanism of two de novo genes, CG31955 and CG32711, in the Drosophila melanogaster species group by knocking out the two genes. Sequence analysis and experimental evidences at both RNA and protein levels demonstrate that both genes are protein-coding and express at multiple developmental stages of D. melanogaster. Targeted mutagenesis shows that both de novo genes have significant phenotypic effects on fertility and lifespan, two key components of adaptive fitness evolution. CG31955 male mutants have decreased fertility, whereas its female mutants show improved fertility and lifespan. Meanwhile, in CG32711 mutants both male and female fertility were significantly decreased and male lifespan was shortened. These data suggestthat CG31955 may inhibit female fertility and lifespan but promote male fertility, while CG32711 are important in fertility of both sexes and also in lifespan of males. Comparative transcriptomic analyses between mutants and wild type suggest that both de novo genes promote male fertility possibly by regulating chitin-related genes and lysozyme genes, CG31955 inhibits female fertility and lifespan possibly by down-regulating egg-related and lysozyme genes, and CG32711 influences male lifespan possibly by down regulating the genes in oxidative phosphorylation pathway. Our results provide solid biological evidence for functional importance of de novo genes.Fruitfly is a traditional model, because of it is easy to breeding and short growth cycle, has been widely applied in biological research, but the problems it can solve in science arelimited. While, the butterflies attract the scientists because of its colorful. Along with the several butterflies genome has been analysis in recent years and the development of gene editing techniques, the butterflies have opportunity to become a promising system to explore the genetic, evolutionary, and developmental mechanisms underlying morphological diversification and speciation. The common yellow swallowtail Papilio machaon, named as the type-species for all butterflies by Linnaeus, has been a long-term focus in the study of mimicry, vision and learning, and pigmentation. CRISPR/Cas9 has emerged as an efficient tool for gene editing across a wide spectrum of organisms including such model insects as fruitfly, silkworm, mosquito etc. Using CRISPR/Cas9 technology, recently we succeeded in editing genes in Asian common swallowtail Papilio xuthus. However, it is not sure if this technology developed in P. xuthus is similarly efficient for other butterfly species. Here,using the similar method, we realized CRISPR/Cas9 gene editing in common yellow swallowtail P. machaon. Our data show that co-injection of sgRNAs targeting Abdominal-B into P. machaon eggs can induce abnormally morphological mutation in 88.5% larva developed from injected eggs with a high frequency of gene disruption up to 98.2%. Real-time quatitative PCR show a very low expression of Abdominal-B gene in abnormally morphological individuals compared with wild types. This work demonstrates that the methodology developed in P. xuthus also can apply to other butterfly species, and further strengths the potentials of butterfly as new models for genetics, evolution and development studiesUnder the artificial selection, how genetic mechanism evolution is always an outstanding question.In 1859, Darwin published the orgin of species, he discuss the domestication in the firstchapter, use the cases of artificial selection to quote the the great power of natural selection. Goats have beendomesticated for a long time, and have a significant impact on development of human society. In the process of domesticated many traits have changed, such as goats have higher hair quality. We use the Realtime-PCR to validation thecopy number variants in domesticated goats and wild goats. We found that the four genes have more copies in domesticated goats. These data provided evidence for the significant changes of genetic materials though the process of domestication.