摘要
提要应用RNeasy animal mini试剂盒抽提大黄鱼total RNA,逆转录合成模板cDNA;同时综合分析从GenBank数据库查询到已报道的转铁蛋白(Tf)基因序列,先设计并合成扩增引物扩增出大黄鱼转铁蛋白部分基因序列,再应用RACE技术,克隆出大黄鱼转铁蛋白全长cDNA基因,全长2461个核苷酸,编码661个氨基酸。应用WCG软件包,采用DNADIST和NEIGHBOR分析方法,构建了转铁蛋白系统进化树。树图显示与传统的分类地位大致相符,可以看出转铁蛋白在海水鱼和淡水鱼的进化上的差异,同时也显示大黄鱼Tf与真鲷的同源性最高。
Transferrin （Tf） is a type of important β -protein for iron transportation, and an important component in biological iron circle. RNeasy Animal Mini Kit was used to obtain total RNA from liver tissue of Pseudosciaena crocea cultured in Xiangshan Bay, Ningbo, China, and special primers were designed for the PCR amplification of Tf gene in the species and compared to the counterpart in Genbank. Full-length cDNA of the Tf was obtained after RACE for being 2461bp long. The entire cDNA was arranged and analyzed with WCG software package having a phylogenetic tree build-up in DNADIST and NEIGHBOR. The Pseudosciaena crocea shared the highest homology with Pagrus major （AY335444）, and the difference between the two is clearly shown in phylogenetic tree.
出处
《海洋与湖沼》
CAS
CSCD
北大核心
2007年第6期563-568,共6页
Oceanologia Et Limnologia Sinica
基金
国家高技术研究发展计划("863")项目
2006AA10A405号
浙江省重大项目
2006C12005号
宁波市重大招标项目
2004C100040号。
关键词
大黄鱼
转铁蛋白
克隆
系统发育
Pseudosciaena crocea, Transferrin （Tf）, Clone, Phylogenic evolution
