CAJ | 학술논문

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1923 bp,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.
위탐토잡교라비어항응격유전능력급기여친본지간관계,채용RT-PCR방법극륭료오니잡교라비어급기친본열휴극단백Hsp70기인완정편마구(code sequences,CDS)적cDNA서렬.서렬분석결과표명:잡교오니라비어급기부본오리아라비어화모본니라라비어열휴극단백Hsp70기인CDS균위1923 bp,편마640개안기산,함유84개감성안기산,95개산성안기산,이론등전점위5.462.통과antheprot분석발현Hsp70가족적3개첨명서렬분별위 IDLGTTYS,IFDLGGGTFD,VVLVGGSTRIPKIQK,핵정위신호표첨:KRKHKKDISQNKRALRR,Dank특정기서DLGTT-S-V,포질Hsp70특정기서EEVD,고근C단적GGMP4태서렬,령유2개당기화위점NKSI화NVSA.오니잡교라비어여기친본Hsp70적서렬비대발현,오니잡교라비어Hsp70적핵감산(100%)화안기산서렬(99.8%)여부본오리아라비어기본일치,여모본니라라비어유2개안기산불동,5개핵감산돌변위점;여모본니라라비어、막상비극라비어、청장、아평안기산서렬동원성분별위99.5%、99.4%、93.1%、83.9%.계통발육수분석표명오니잡교라비어여부본오리아라비어거리최근,여모본니라라비어거리원,해결과여전통잡교라비어편부본유전결론상부.
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） Hsp 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: IDLGTTYS，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 C-extreme 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.