【摘要】：銜接蛋白2(adaptin protein 2,Ap2)是披網格蛋白小泡(clathrin-coated vesicles,CCVS)的重要組成部分,在蛋白質通過細胞膜進入胞質的轉運過程中發(fā)揮著重要作用。我們通過人類睪丸cDNA芯片的方法將成人和6月齡的胚胎睪丸進行雜交比較,克隆了一個銜接蛋白2β(Ap2β)亞單位的不同剪接體-Ap2β-NY。芯片雜交結果顯示,Ap2β-NY在成人睪丸中mRNA的表達量是胚胎睪丸的3.62倍。對Ap2β-NY基因進行測序,獲得全長序列并輸入GenBank,登錄號為AY341427。生物學軟件分析表明,全長為3457bp的Ap2β-NY基因編碼了一個具有880個氨基酸的蛋白質,預測此蛋白質的分子量為98.12KDa,等電點PI為5.01,定位于人17號染色體上的q11.2-q12。對預測蛋白進行生物信息學分析發(fā)現,Ap2β-NY具有Ap2β的全部三個結構域,即具有一個Ap2β氮末端結構域,一個低度重復序列和一個Ap2β炭末端結構域。但是Ap2β-NY的氮末端結構域比Ap2β短了57個氨基酸殘基,并且這57個氨基酸殘基是疏水性的,很有可能位于蛋白質的表面,是蛋白質相互作用的結合位點。用反轉錄PCR(reverse transcription PCR,RT-PCR)的方法檢測成人和胚胎睪丸Ap2β-NY的表達狀況,證實了芯片雜交結果。同時精子RT-PCR表明Ap2β-NY在精子中也高表達。用Southern雜交的方法檢測Ap2β-NY在16種正常人體組織中的表達情況,發(fā)現Ap2β-NY高表達于脾臟、胸腺、前列腺、卵
[Abstract]:Binding protein 2 (adaptin protein 2 / Ap2 is an important component of drape grid protein vesicles (clathrin-coated vesicles,CCVS), which plays an important role in the transport of proteins into the cytoplasm through the cell membrane. Using human testis cDNA microarray, we compared adult testis with 6-month-old embryonic testis and cloned Ap2 尾 NY., a splice of different splicing units of Ap2 尾. The results of microarray hybridization showed that the expression of Ap2 尾-NY in adult testis was 3.62 times higher than that in embryonic testis. The Ap2 尾-NY gene was sequenced, the full-length sequence was obtained and the GenBank, login number was entered as AY341427.. The analysis of biological software shows that the Ap2 尾-NY gene with full length of 3457bp encodes a protein with 880 amino acids. The predicted molecular weight of the protein is 98.12 KDaand the isoelectric point PI is 5.01, which is located at q11.2-q12 on human chromosome 17. Bioinformatics analysis showed that Ap2 尾-NY had all three domains of Ap2 尾, namely, one Ap2 尾 N-terminal domain, one low-degree repeat sequence and one Ap2 尾 C-terminal domain. However, the nitrogen terminal domain of Ap2 尾-NY is 57 amino acid residues shorter than Ap2 尾, and the 57 amino acid residues are hydrophobic and probably located on the surface of protein and are binding sites for protein interaction. Reverse transcription PCR (reverse transcription PCR,RT-PCR (RT PCR (reverse transcription PCR,RT-PCR) was used to detect the expression of Ap2 尾-NY in adult and embryonic testis, which confirmed the results of microarray hybridization. At the same time, sperm RT-PCR showed that Ap2 尾-NY was also highly expressed in spermatozoa. The expression of Ap2 尾-NY in 16 normal human tissues was detected by Southern hybridization. It was found that Ap2 尾-NY was highly expressed in spleen, thymus, prostate and egg.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2005
【分類號】：R341

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