Noguchi Yoko, Takano Kaoru, Koura Minako, Uchio-Yamada Kozue, Matsuda Junichiro, Suzuki Osamu
Laboratory of Experimental Animal Models, National Institute of Biomedical Innovation, Saito-Asagi 7-6-8, Ibaraki, Osaka 567-0085, Japan.
Gen Comp Endocrinol. 2006 Jun;147(2):231-5. doi: 10.1016/j.ygcen.2006.01.001. Epub 2006 Feb 14.
To understand the molecular basis of the rabbit's efficient superovulation, we determined the cDNA sequence of the follicle-stimulating hormone (FSH) beta-subunit precursor protein using a combination of 5'- and 3'-rapid amplification of cDNA ends (RACE) with pituitary cDNA libraries of the Japanese White rabbit and compared it with those of other mammals. RACE experiments detected at least three transcripts for the FSHbeta precursor protein in the libraries. The transcripts had lengths of 457, 1,621, and 1,767 bp, from the 5'-end to the poly(A) site. The shortest and mid-length transcripts had the putative polyadenylation signal sequence AATAAA at nucleotides 436 and 1,601, respectively, whereas the longest form had an ATTAAA sequence at nucleotide 1,745 of the cDNA sequence. These transcripts are likely to be polyadenylation variants of one large transcript because they share the same coding sequence for the precursor protein (130 amino acid residues in length). However, only a few shortest variants seem to be formed because the shortest variants were not detected by Northern blot analysis. Phylogenetic analysis of the deduced amino acid sequence indicates that the rabbit is phylogenetically closer to humans than to the other mammals, suggesting that an FSH preparation from human sources would be superior as a follicle stimulant for the induction of superovulation.
为了解兔子高效超排卵的分子基础，我们结合5'-和3'-cDNA末端快速扩增（RACE）技术，利用日本白兔垂体cDNA文库测定了促卵泡激素（FSH）β亚基前体蛋白的cDNA序列，并将其与其他哺乳动物的进行比较。RACE实验在文库中检测到至少三种FSHβ前体蛋白的转录本。这些转录本从5'-末端到poly(A)位点的长度分别为457、1621和1767 bp。最短和中等长度的转录本在核苷酸436和1601处分别具有推定的聚腺苷酸化信号序列AATAAA，而最长形式在cDNA序列的核苷酸1745处具有ATTAAA序列。这些转录本可能是一个大转录本的聚腺苷酸化变体，因为它们共享前体蛋白相同的编码序列（长度为130个氨基酸残基）。然而，似乎只形成了少数最短的变体，因为最短的变体在Northern印迹分析中未被检测到。对推导氨基酸序列的系统发育分析表明，兔子在系统发育上比其他哺乳动物更接近人类，这表明来自人类来源的FSH制剂作为诱导超排卵的卵泡刺激剂可能更优越。
