CAJ | 학술논문

目的筛选新的卵巢特异性蛋白质,探讨其可能在卵细胞成熟及胚胎发育过程中作用.方法收集小鼠成熟卵细胞,经二维电泳、蛋白印迹、质谱鉴定,获得新的多肽序列;应用RT、RACE-PCR等技术,获得该段多肽对应的新蛋白的全长cDNA序列,并行Northernblot分析其组织特异性.结果所获得的多肽序列来自一种未知新蛋白,其对应的cDNA全长有1510 bp,前32 bp为5'非编码区,第33～1433 bp为ORF区,第1434～1510 bp为3'非编码区;该基因定位于9号染色体上,预测表达466个氨基酸,理论相对分子质量和pI分别为53 400和8.05,具有稳定WD40,F-box保守性功能域,并特异性表达于卵巢组织.结论一种具有卵巢组织特异性的新蛋白被发现,其生物信息学特征表明其可能参与卵细胞内分子信号通道,可能在卵成熟及早期胚胎发育中具有作用.
목적 사선신적란소특이성단백질,탐토기가능재란세포성숙급배태발육과정중작용.방법 수집소서성숙란세포,경이유전영、단백인적、질보감정,획득신적다태서렬;응용RT、RACE-PCR등기술,획득해단다태대응적신단백적전장cDNA서렬,병행Northernblot분석기조직특이성.결과 소획득적다태서렬래자일충미지신단백,기대응적cDNA전장유1510 bp,전32 bp위5'비편마구,제33～1433 bp위ORF구,제1434～1510 bp위3'비편마구;해기인정위우9호염색체상,예측표체466개안기산,이론상대분자질량화pI분별위53 400화8.05,구유은정WD40,F-box보수성공능역,병특이성표체우란소조직.결론 일충구유란소조직특이성적신단백피발현,기생물신식학특정표명기가능삼여란세포내분자신호통도,가능재란성숙급조기배태발육중구유작용.
Objective To screen and identify the novel ovarian specific proteins to explore the possible mechanisms in the oogenesis and early embryo development. Methods The oocytes of BALB/C mice were collected for 2-D electrophoresis. After reaction with anti-citrulline antibody, three positive spots were selected for mass spectrometry. Based on the biologic informatic analysis, a novel peptide sequence was found and specific primers were designed for cloning. A full-length cDNA was identified by reverse transcription and rapid amplification of cDNA ends - polymerase chain reaction ( RACE-PCR ) . Its nucleotide and possible protein sequences were compared with those in the gene bank. And the tissuespecific characteristic of MOP53 was analyzed by Northern blot. Results A novel peptide sequence was found in murine oocytes, it came from a predicted protein with 330 amino acids. The predicted protein had ～53 400 and pI 8. 05 with conserved WD40, F-box motif. A full-length cDNA sequence with 1510 bp was cloned and named MOP53. Located at chromosome 9 F2, this new gene was specifically expressed in ovary.Conclusions An ovarian specific protein is found. It may be involved in the process of oogenesis and early embryo development based on its biologic informatic characteristics.