CAJ | 학술논문

利用RACE技术首次克隆了半滑舌鳎脑垂体中促黄体激素（LH）基因cDNA全长序列。该基因全长670 bp，开放阅读框为477 bp，编码158个氨基酸。与其他脊椎动物的LH成熟肽氨基酸序列同源性比较表明：半滑舌鳎LH与鲽形目和鲈形目同源性高58%～68%。另外，半滑舌鳎LH含有12个保守的半胱氨酸残基和1个N-糖基化位点（19～21 NQT）。实时荧光定量组织表达分析表明，LH mRNA在脑、垂体、卵巢等组织中表达，垂体中表达量最丰富，其他组织尤以脑、性腺表达量较高，推测半滑舌鳎LH可能具有广泛的垂体外生理功能；实时荧光定量PCR方法对繁殖周期雌性半滑舌鳎LH表达水平进行测定，结果表明， LH mRNA在Ⅱ～Ⅵ各繁殖周期的脑、垂体、卵巢3种组织都有表达，但表达水平有差异，在Ⅳ、Ⅴ期表达量最丰富，说明LH主要促进卵母细胞最终成熟及排卵。利用125I标记的放射免疫测定（RIA）技术检测繁殖周期半滑舌鳎血清LH浓度，结果表明LH在Ⅴ期含量最丰富，也说明LH主要促进卵母细胞最终成熟及排卵。
이용RACE기술수차극륭료반활설탑뇌수체중촉황체격소（LH）기인cDNA전장서렬。해기인전장670 bp，개방열독광위477 bp，편마158개안기산。여기타척추동물적LH성숙태안기산서렬동원성비교표명：반활설탑LH여접형목화로형목동원성고58%～68%。령외，반활설탑LH함유12개보수적반광안산잔기화1개N-당기화위점（19～21 NQT）。실시형광정량조직표체분석표명，LH mRNA재뇌、수체、란소등조직중표체，수체중표체량최봉부，기타조직우이뇌、성선표체량교고，추측반활설탑LH가능구유엄범적수체외생리공능；실시형광정량PCR방법대번식주기자성반활설탑LH표체수평진행측정，결과표명， LH mRNA재Ⅱ～Ⅵ각번식주기적뇌、수체、란소3충조직도유표체，단표체수평유차이，재Ⅳ、Ⅴ기표체량최봉부，설명LH주요촉진란모세포최종성숙급배란。이용125I표기적방사면역측정（RIA）기술검측번식주기반활설탑혈청LH농도，결과표명LH재Ⅴ기함량최봉부，야설명LH주요촉진란모세포최종성숙급배란。
The full length cDNA of LH has been cloned from pituitary of half smooth tongue sole C. Günther by using RACE technology for the first time. The cDNA of LH is 670 bp,and the open reading frame is 477 bp encoding peptides of 158 amino acid. Multiple amino acid se-quences alignments were performed among vertebrates,and the results showed that tongue sole LH mature peptide was the most similar to their orthologs of Pleuronectiformes and Perciformes which was 58%~68%identity. And 12 conserved cysteine residues and 1 N-linked glycosyl-ation site were found in the deduced amino acid sequence. Real-time PCR analysis showed that LH mRNA was expressed in all twelve tissues including brain,pituitary,ovary and so on. The highest expression level was in pituitary. And the stronger amplification signals were detected in the brain and gonad than all other non-pituitary tissues. It suggested that LH may had the more physiological functions because LH mRNA of tongue sole expressed widely in non-pitu-itary tissues. Quantitative real-time PCR was used for analyzing the expression of female C. semilaevis LH mRNA in the reproductive cycle. The results showed that LH mRNA was ex-pressed in all three tissues including brain,pituitary and ovary from the stage Ⅱto stage Ⅵ, and the highest expression level appeared on the stageⅣ and stageⅤwhich suggested that the main function of LH is to promote eggs finally mature and discharge. Radio immunoassay (RIA)marked by 125I was used to detect the concentration of plasma LH in C.semilaevis. The re-sults showed that the biggest plasma LH concentration appareed on the stage of Ⅴwhich also suggested that the main function of LH was to promote eggs finally mature and discharge.