CAJ | 학술논문

采用同源性克隆和cDNA末端快速扩增（RACE）方法，从条石鲷（Oplegnathidae f asciatus）垂体中获得了全长为897 bp的GtHαcDNA序列，其中包含249 bp的5′非编码区、399 bp的开放阅读框和249 bp的3′非编码区。该基因编码由132个氨基酸组成的前体蛋白，其中前38个氨基酸为信号肽，在第122和124Cys中间间隔了一个组氨酸（His），形成CXC型趋化因子的特征结构，肽链中包括10个保守Cys残基和2个N‐糖基化位点。氨基酸同源性和进化分析表明，条石鲷GtHα与鲈形目鱼类的进化关系较近，与鲈形目鱼类 GtHα的同源性为87％～98％，与其他脊椎动物GtHα的同源性为53％～87％。实时荧光定量PCR检测发现，GtHαmRNA在垂体、脑、性腺中的表达量较高，在心、肝脏、胃、肠、肌肉、幽门盲囊中微量表达，在脾脏、头肾、鳃、肾脏中不表达，其中在垂体中的表达量最高；周期表达发现：垂体中GtHαmRNA在卵巢发育的Ⅴ期达到最大值，卵巢中GtHαmRNA在卵巢发育的Ⅳ期达到最大值，而脑组织中GtHαmRNA在卵巢发育的Ⅴ期达到最低值，表明GtHα对卵母细胞发育的调控可能同时通过自分泌和旁分泌两种途径进行。本研究为条石鲷生殖调控机制及人工繁育技术研究提供了基础资料。
채용동원성극륭화cDNA말단쾌속확증（RACE）방법，종조석조（Oplegnathidae f asciatus）수체중획득료전장위897 bp적GtHαcDNA서렬，기중포함249 bp적5′비편마구、399 bp적개방열독광화249 bp적3′비편마구。해기인편마유132개안기산조성적전체단백，기중전38개안기산위신호태，재제122화124Cys중간간격료일개조안산（His），형성CXC형추화인자적특정결구，태련중포괄10개보수Cys잔기화2개N‐당기화위점。안기산동원성화진화분석표명，조석조GtHα여로형목어류적진화관계교근，여로형목어류 GtHα적동원성위87％～98％，여기타척추동물GtHα적동원성위53％～87％。실시형광정량PCR검측발현，GtHαmRNA재수체、뇌、성선중적표체량교고，재심、간장、위、장、기육、유문맹낭중미량표체，재비장、두신、새、신장중불표체，기중재수체중적표체량최고；주기표체발현：수체중GtHαmRNA재란소발육적Ⅴ기체도최대치，란소중GtHαmRNA재란소발육적Ⅳ기체도최대치，이뇌조직중GtHαmRNA재란소발육적Ⅴ기체도최저치，표명GtHα대란모세포발육적조공가능동시통과자분비화방분비량충도경진행。본연구위조석조생식조공궤제급인공번육기술연구제공료기출자료。
A full length cDNA sequence of gonadotropin hormone common alpha subunit was cloned from the pituitary of Oplegnathus fasciatususing by homology cloning and RACE techniques .The GtHα cDNA sequence consisted of 897 base pairs nucleotides ,including 249 base pairs at 5′untranslation region ,249 base pairs at 3′untranslation region and an open reading frame with 399 base pairs nucleotides .The cDNA sequence encoded a precursor protein of 132 amino acids preceded by a signal peptide of 38 amino acids resi‐dues .T he putative peptide contained ten cysteine residues and tw o N‐glycosylation sites residues .A histi‐dine residue in the middle of 122 and 124 cysteine formed a CXC chemokine characteristic structure .Se‐quences comparison and phylogenetic analysis showed that the GtHαof Oplegnathus f asciatus had higher identity to their orthologs of Perciformes (87% ~98% ) than to other vertebrates (53% ~87% ) .The qRT‐PCR results suggested that the level of GtHα mRNA was higher in pituitary ,ovary ,brain and trace ex‐pression in heart ,liver ,stomach ,intestine ,muscle and pyloric caecum .However ,it was not found in kid‐ney ,Head kidney ,spleen and gill by Real‐time Quantitative PCR technique .The level of GtHα mRNA was highest in pituitary .The reproductive cycle expression showed that the level of GtHαmRNA in pitui‐tary was maximum in the stage Ⅴ of ovarian development and that in ovary was maximum in the stage Ⅳ . However ,the level in brain was lowest in the stage Ⅴ suggesting the regulation of GtHαin the ovarian de‐velopment may through both autocrine and paracrine pathways .This research provides a reference to the reproductive regulation mechanism and artificial breeding technigues of Oplegnathus f asciatus .