An efficient microinjection method for unfertilized eggs of Asian amphioxus Branchiostoma belcheri

CORC > 厦门大学 > 海洋环境－已发表论文

	An efficient microinjection method for unfertilized eggs of Asian amphioxus Branchiostoma belcheri
	Liu, Xin ; Li, Guang ; Feng, Jun ; Yang, Xi ; Wang, Yi-Quan ; Liu X(柳欣) ; Li G(李光) ; Wang YQ(王义权)
刊名	http://dx.doi.org/10.1007/s00427-013-0441-0
	2013
关键词	GREEN FLUORESCENT PROTEINS EMBRYOS EVOLUTIONARY CULTURE BIOLOGY GFP
英文摘要	National High Technology Research and Development Program of China [2008AA092602]; National Natural Science Foundation of China [30830023, 31101631]; Amphioxus is a promising model animal for evolutionary and developmental studies. However, as an emerging model organism, amphioxus lacks most molecular techniques applied in other well-developed model animals. Microinjection is a powerful technique for gene manipulation, and thus it undoubtedly is one of useful approaches in the studies of gene function and embryonic development. Although the method has been exploited in Florida and European amphioxus, it still remains to be optimized and introduced into other amphioxus species. In order to introduce the technique into our lab, we followed and optimized the previous description and successfully performed microinjection on unfertilized eggs of Asian amphioxus Branchiostoma belcheri. We made six solutions for practice: 200 mM KCl, 100 ng/mu l actin-LacZ or 100 ng/mu l actin-RFP vectors, LacZ or RFP vectors without promoter and RFP capped mRNA. More than 99.2 % of eggs injected with KCl were able to be fertilized, 94.3 % of them could hatch normally and 55.9 % survived until 2-day larvae, all of which were nearly equivalent to those obtained from normally fertilized eggs. Embryos injected with two plasmid constructs also showed very high fertilizing and hatching ratios, but normally developing ratios were slightly lower than that of KCl injection. Of those injected embryos, 91.8 % expressed exogenous gene LacZ and 80.5 % exhibited foreign RFP expression, which were driven by a promoter from amphioxus beta-actin gene. The data indicated a successful modified microinjection method for the unfertilized eggs of Asian amphioxus, and those modifications improved the feasibility and efficiency of microinjection on amphioxus.
语种	英语
出版者	SPRINGER
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/88003]
专题	海洋环境－已发表论文
推荐引用方式 GB/T 7714	Liu, Xin,Li, Guang,Feng, Jun,et al. An efficient microinjection method for unfertilized eggs of Asian amphioxus Branchiostoma belcheri[J]. http://dx.doi.org/10.1007/s00427-013-0441-0,2013.
APA	Liu, Xin.,Li, Guang.,Feng, Jun.,Yang, Xi.,Wang, Yi-Quan.,...&王义权.(2013).An efficient microinjection method for unfertilized eggs of Asian amphioxus Branchiostoma belcheri.http://dx.doi.org/10.1007/s00427-013-0441-0.
MLA	Liu, Xin,et al."An efficient microinjection method for unfertilized eggs of Asian amphioxus Branchiostoma belcheri".http://dx.doi.org/10.1007/s00427-013-0441-0 (2013).