摘要
为探讨杂交罗非鱼抗应激遗传能力及其与亲本之间关系,采用RT-PCR方法克隆了奥尼杂交罗非鱼及其亲本热休克蛋白Hsp70基因完整编码区(code sequences,CDS)的cDNA序列。序列分析结果表明:杂交奥尼罗非鱼及其父本奥利亚罗非鱼和母本尼罗罗非鱼热休克蛋白Hsp70基因CDS均为1923bp,编码640个氨基酸,含有84个碱性氨基酸,95个酸性氨基酸,理论等电点为5.462。通过antheprot分析发现Hsp70家族的3个签名序列分别为IDLGTTYS,IFDLGGGTFD,VVLVGGSTRIPKIQK,核定位信号标签:KRKHKKDISQNKRALRR,Dank特征基序DLGTT-S-V,胞质Hsp70特征基序EEVD,靠近C端的GGMP4肽序列,另有2个糖基化位点NKSI和NVSA。奥尼杂交罗非鱼与其亲本Hsp70的序列比对发现,奥尼杂交罗非鱼Hsp70的核苷酸(100%)和氨基酸序列(99.8%)与父本奥利亚罗非鱼基本一致,与母本尼罗罗非鱼有2个氨基酸不同,5个核苷酸突变位点;与母本尼罗罗非鱼、莫桑比克罗非鱼、青鳉、牙鲆氨基酸序列同源性分别为99.5%、99.4%、93.1%、83.9%。系统发育树分析表明奥尼杂交罗非鱼与父本奥利亚罗非鱼距离最近,与母本尼罗罗非鱼距离远,该结果与传统杂交罗非鱼偏父本遗传结论相符。
Hybrid is the main and general method for getting high male of tilapia at present. In this study, we designed to find the relationships of anti-stress between Oreochromis niloticus ♀× Oreochromis aureus ♂ and their parents. The code sequences of heat shock protein 70 gene) lisp 70 ) were amplied and cloned from O. niloticus ♀× O. aureus ♂ and their parents by RT-PCR. The sequence analysis showed that their CDS sequences were 1 923 bp,encoding 640 amino acids with 84 strongly basic ( + ) ,95 strongly acidic ( - ) and isolectric point of 5. 462. The CDS have three signature sequences of Hsp 70 family: IDLGrITYS,IFDLGGGTFD and VVLVGGSTRIPKIQK, the sequence of bipartite nuclear targeting KRKHKKDISQNKRALRR, the sequence of Dank subfamily DLGTY-S-V, the sequence of cytoplasm EEVD, four peptide sequence GGMP closely Cextreme and two glycosylation site NKSI and NVSA by antheprot. The analysis also revealed the Hsp70 sequence of O. niloticus ♀× O. aureus ♂ was similar to O. aureus in nucleotides( 100% ) and amino acids(99.8% ), but different in 2 amino acids and 5 nucleotides to O. niloticus. The amino acids sequence of O. niloticus ♀× O. aureus ♂ showed 99.5% ,99.4% ,93.1% ,83.9% identity with O, niloticus,O. mossambicus, Oryzias latipes and Paralichthys olivaceus, respectively. The results of phylogenetic analysis indicated that the Hsp70 sequence of O. niloticus ♀× O. aureus ♂ was very close to O. aureus and far away from O. niloticus. It was consistent with the O. niloticus ♀× O. aureus ♂ inheriting from O. aureu.
出处
《上海海洋大学学报》
CAS
CSCD
北大核心
2009年第4期391-397,共7页
Journal of Shanghai Ocean University
基金
广西自然科学资金(No.0731036)
公益性行业(农业)科研专项(3-49)
关键词
罗非鱼
HSP70基因
基因克隆
序列分析
Oreochromis niloticus ♀× Oreochromis aureus ♂
Hsp70 gene
clone
sequence analysis