Isolation of cDNA for a Xenopus sperm-specific basic nuclear protein (SP4) and evidence for expression of SP4 mRNA in primary spermatocytes.
Isolation of cDNA for a Xenopus sperm-specific basic nuclear protein (SP4) and evidence for expression of SP4 mRNA in primary spermatocytes.
复制标题
非洲爪蟾精子特异性碱性核蛋白 (SP4) cDNA 的分离以及原代精母细胞中 SP4 mRNA 表达的证据。
DOI：
10.1016/0014-4827(91)90135-h
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发表时间：
1991
影响因子：
3.7
通讯作者：
S. Abe
中科院分区：
文献类型：
--
作者：
H. Hiyoshi;S. Uno;T. Yokota;C. Katagiri;H. Nishida;M. Takai;K. Agata;G. Eguchi;S. Abe
从Xenopus圆形精子细胞中分离poly（A）+mRNA，用抗精子特异蛋白SP3-5抗体筛选，在λ gt 11中构建cDNA文库。对阳性克隆进行测序，并获得了一个富含精氨酸的克隆，命名为pXSP 531。pXSP 531全长473个核苷酸，包含一个237个核苷酸的开放阅读框，其前有一个67个核苷酸的5′端非翻译区。3′端非翻译区包含149个核苷酸，其中包括一个多聚腺苷酸化信号（AAATAAAA）。pXSP 531中含有20个聚腺苷酸尾核苷酸。SP3-5通过反相色谱分离并测序。通过用V8蛋白酶消化SP 4并通过反相色谱分离获得的肽片段的氨基酸序列与从pXSP 531的核苷酸序列推导的N-末端43和C-末端15个氨基酸的序列相同。该结果表明pXSP 531编码SP 4。用SP 4 cDNA探针（pXSP 531）对第0天和第6天从初级精母细胞和圆形精子细胞中提取的RNA进行北方杂交，结果表明，SP 4 mRNA在初级精母细胞和圆形精子细胞中均存在，虹鳟鱼的鱼精蛋白mRNA也存在。第0天圆形精子细胞中SP 4 mRNA的大小比初级精母细胞长60个核苷酸，第6天精子细胞中SP 4 mRNA的大小比第0天短30个核苷酸。这些大小差异是由于poly（A）片段长度的差异，因为用核糖核酸酶H消化poly（A）导致mRNA缩短到相同大小的三个阶段。
A cDNA library was prepared in λgt11 from poly(A)+mRNA isolated from a pure population ofXenopusround spermatids and screened with an antibody against SP3–5 (sperm-specific proteins) ofXenopussperm. Positive clones were sequenced and an arginine-rich clone, designated pXSP531, was obtained. The 473-nucleotide sequence of pXSP531 contained an open reading frame of 237 nucleotides which was preceded by a 5′ untranslated region of 67 nucleotides. The 3′ untranslated region contained 149 nucleotides, including a consensus polyadenylation signal (AAATAAAA). Twenty nucleotides of a poly(A) tail was contained in the pXSP531. SP3–5 were separated from each other by reverse-phase chromatography and sequenced. The amino acid sequence of the peptide fragments which were obtained by digestion of SP4 with V8 protease and separated by reverse-phase chromatography was identical to the sequence of the N-terminal 43 and C-terminal 15 amino acids deduced from the nucleotide sequence of pXSP531. This result demonstrates that pXSP531 encodes SP4. Northern hybridization of RNA extracted from primary spermatocytes and round spermatids on Days 0 and 6 with SP4 cDNA probe (pXSP531) showed that SP4 mRNA is present both in primary spermatocytes and in round spermatids as is protamine mRNA in the rainbow trout. The size of the SP4 mRNA in round spermatids on Day 0 was longer by 60 nucleotides compared to that in primary spermatocytes and that in spermatids on Day 6 was shorter by 30 nucleotides compared to that on Day 0. These size differences were due to differences in the length of the poly(A) tracts because digestion of poly(A) with ribonuclease H resulted in the shortening of mRNA to the same size for three stages.
影响因子：
3.7
作者：
HECHT, NB;BOWER, PA;DISTEL, RJ
通讯作者：
DISTEL, RJ
DOI：
10.1016/s0021-9258(18)45202-7
发表时间：
1987-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. Heidaran;W. Kistler
通讯作者：
M. Heidaran;W. Kistler
鱼精蛋白 1 mRNA 的大小变化为监测野生型和突变型小鼠的精子发生提供了分子标记。
DOI：
10.1111/j.1432-0436.1985.tb00314.x
发表时间：
1985
期刊：
Differentiation; research in biological diversity
影响因子：
--
作者：
Hecht,NB;Bower,PA;Kleene,KC;Distel,RJ
通讯作者：
Distel,RJ