CAJ | 학술논문

通过RT-PCR、3'-RACE、5'-RACE方法,从6种重要经济鱼类--大眼鳜(Siniperca kneri)、石斑鱼(Epinephelus coioides)、黄鳝(Monopterus albus)、鲶鱼(Silurus asotus)、泥鳅(Misgurnus anguillicaudatus)和方正银鲫(Carassius auratus gibelio Bloch,Fang Zheng crucian carp)中克隆了生长激素(Growth Hormone,GH)的完整cDNA序列(除石斑鱼序列外,其他生长激素序列均系第一次克隆),并详细分析了其序列特征.测序结果显示,克隆的6种GH cDNA长度依次为953 bp、1 023 bp、825 bp、1 082 bp、1 154 bp和1 180 bp,它们均包含一个长度为600个左右核苷酸的完整阅读框,分别编码一个200个左右氨基酸的蛋白:大眼鳜、石斑鱼和黄鳝GH为204个氨基酸,鲶鱼GH为200个氨基酸,泥鳅和方正银鲫GH为210个氨基酸.这6种蛋白序列与其他已知的鱼类GH序列都有较高的同源性,特别是与相同目的鱼类序列相比.通过序列比对,在这些蛋白序列内鉴定了许多保守的氨基酸残基,其中的大多数聚集而成5个保守域.基于这6种鱼类序列的编码区和其他鱼类的GH编码序列进行分子系统学分析,结果(MP和NJ树)与根据形态特征构建的系统发育树基本一致,特别是在硬骨鱼类较大分类阶元(目间、目以上)的系统发育研究方面比较一致,尽管仍存在一定差异,说明生长激素基因的编码区应该在硬骨鱼类系统发育研究领域得到更多的重视.
통과RT-PCR、3'-RACE、5'-RACE방법,종6충중요경제어류--대안궐(Siniperca kneri)、석반어(Epinephelus coioides)、황선(Monopterus albus)、염어(Silurus asotus)、니추(Misgurnus anguillicaudatus)화방정은즉(Carassius auratus gibelio Bloch,Fang Zheng crucian carp)중극륭료생장격소(Growth Hormone,GH)적완정cDNA서렬(제석반어서렬외,기타생장격소서렬균계제일차극륭),병상세분석료기서렬특정.측서결과현시,극륭적6충GH cDNA장도의차위953 bp、1 023 bp、825 bp、1 082 bp、1 154 bp화1 180 bp,타문균포함일개장도위600개좌우핵감산적완정열독광,분별편마일개200개좌우안기산적단백:대안궐、석반어화황선GH위204개안기산,염어GH위200개안기산,니추화방정은즉GH위210개안기산.저6충단백서렬여기타이지적어류GH서렬도유교고적동원성,특별시여상동목적어류서렬상비.통과서렬비대,재저사단백서렬내감정료허다보수적안기산잔기,기중적대다수취집이성5개보수역.기우저6충어류서렬적편마구화기타어류적GH편마서렬진행분자계통학분석,결과(MP화NJ수)여근거형태특정구건적계통발육수기본일치,특별시재경골어류교대분류계원(목간、목이상)적계통발육연구방면비교일치,진관잉존재일정차이,설명생장격소기인적편마구응해재경골어류계통발육연구영역득도경다적중시.
In this study,the full-length cDNAs of GH (Growth Hormone) gene was isolated from six important economic fishes,Siniperca kneri,Epinephelus coioides,Monopterus albus,Silurus asotus,Misgurnus anguillicaudatus and Carassius auratus gibelio Bloch.It is the first time to clone these GH sequences except E.coioides GH.The lengths of the above cDNAs are as follows: 953 bp,1 023 bp,825 bp,1 082 bp,1 154 bp and 1 180 bp.Each sequence includes an ORF of about 600 bp which encodes a protein of about 200 amino acid: S.kneri,E.coioides and M.albus GHs of 204 amino acid,S.asotus GH of 200 amino acid,M.anguillicaudatus and C.auratus gibelio GHs of 210 amino acid.Then detailed sequence analysis of the six GHs with many other fish sequences was performed.The six sequences all showed high homology to other sequences,especially to sequences within the same order,and many conserved residues were identified,most localized in five domains.The phylogenetic trees (MP and NJ) of many fish GH ORF sequences (including the new six) with Amia calva as outgroup were generally resolved and largely congruent with the morphology-based tree though some incongruities were observed,suggesting GH ORF should be paid more attention to in teleostean phylogeny.