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显微注射技术在制备鱼类嵌合体和转基因海水鱼上的应用   总被引:1,自引:0,他引:1  
以鱼类胚胎细胞和胚胎干细胞为核供体进行细胞移植构建鱼类嵌合体研究方面,现有的成功报道均采用显微注射方法;在转基因海水鱼类研究中,显微注射也是最为常用的技术,本实验室在花鲈胚胎干细胞嵌合体构建和外源基因向花鲈胚胎的转移研究中取得的结果也充分证实,显微注射技术是开展海水鱼类细胞移植和转基因研究的首选技术。  相似文献   
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绿色荧光蛋白基因向花鲈胚胎的转移及其表达   总被引:3,自引:0,他引:3  
通过显微注射的方法,将绿色荧光蛋白(Green fluorescent protein,GFP)基因注射入花鲈(Lateolabrax japonicus)单细胞期受精卵动物极细胞质内。初步研究了显微注射后的花鲈胚胎的存活状况。在荧光显微镜下观察到了GFP的表达,多数胚胎表现为嵌合性表达。利用PCR技术,从花鲈尾芽期胚胎DNA中扩增出了特异片段,大小为750bp,表明显微注射胚胎中整合了外源GFP基因。  相似文献   
3.
真鲷天然抗性相关巨噬蛋白全长cDNA的克隆与序列分析   总被引:1,自引:0,他引:1  
徐美瑜 《水产学报》2005,29(1):128-132
Natural resistance associated macrophage protein (Nmmp) is an innate resistance protein to intracellular parasites, which is expressed plentifully in macrophage ceils. Nramp has been studied in mouse, human, cattle, rainbow trout and channel catfish.However, tittle was known about the structure of Pagrus major Nramp. In order to get the complete sequence of Pagrus major Nramp, a pair of primer is designed according to a 200bp known sequence of Pagrus major Nramp cDNA. By the use of SMART RACE, the full Nramp of Pagrus major cDNA about 5 000 bp was obtained, including about 200 bp 5‘ terminal region (UTR),complete encoding region and 3‘ terminal region. There were 3 ployA signals, which showed many possibilities of cutting at 3‘ terminal region. The character of Pagrus major Nramp nucleotide sequence and deduced amino acid sequence are analyzed. 12 putative transmembrane(TM) regions, a consensus transport motif (CTM), a predicted protein kinase C phosphroylation site and three predicted N-link glycosylation sites are indicated in its deduced amino acid sequence. The ‘consense transport motif‘ CTM is located etween TM8 and TM9. Furthermore, a protein kinase C phosphroylation site and three N-link glycosylation sites were predicted. The lignment of amino acid sequences between Pagrus major Nramp cDNA and several animals is analyzed and the deduced amino acid equence of Pagrus major Nrarnp had 77.8%, 83.0%, 82.3%, 80.0%, 81.1%, 60.4%, 70.3%, 58.5%, 69.5% identity ith rainbow trout α(AAD20721), rainbow trout β(AAD20722), channel catfish(AF400108), fathead minnow (AAF01778),common carp (CABal96), mouse 1 ( AAA39838 ), mouse 2 ( AAC42051 ), human 1 ( D50403 ), human 2 ( NP - 0(106(18 ),respectively. The alignment reveals high conservation in TM and CTM regions. Analysis result makes us get familiar with the structure nd character of fish Nramp, furthermore, offers some infonnat/on for the enhancement of immunity of fish and genetic amelioration on fish breeding.  相似文献   
4.
鱼类基因转移技术研究进展   总被引:5,自引:2,他引:3  
基因转移为鱼类遗传育种开辟了一条新的途径。自1985年第1例转基因鱼问世至今已获得十几种转基因鱼,在促进生长,提高鱼类抗逆性、抗病性等方面取得了巨大的成绩。本文主要结合国内外关于鱼类基因转移成功的报道,分析了各种转基因技术的可行性和优缺点,并对其作用机理、适用对象、条件优化、转化效率等几个方面做了概括性叙述。  相似文献   
5.
适合花鲈的几种染色体制备方法的比较   总被引:10,自引:2,他引:8  
比较了6种适于制备花鲈(Lateolabrax japonicus)染色体的制片方法。结果表明,细胞系(LJES)培养法和头肾-PHA活体注射-淋巴分离液富集法可制备高质量的染色体标本;外周血短期培养法和头肾-PHA活体注射法得到的结果重复性好,但制片时有红细胞的干扰;小鱼游泳法和组织浸泡法操作简单,适于野外条件下的染色体制备,但得到的染色体图象有背景杂质,分裂指数较低,仅适用于染色体组型的研究。  相似文献   
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