Rat transition nuclear protein 2 regulatory region directs haploid expression of reporter gene in male germ cells of transgenic mice
Identifieur interne : 000998 ( Main/Exploration ); précédent : 000997; suivant : 000999Rat transition nuclear protein 2 regulatory region directs haploid expression of reporter gene in male germ cells of transgenic mice
Auteurs : Karim Nayernia [Allemagne] ; Detlef Böhm [Allemagne] ; Özlem Topaloglu [Allemagne] ; Gregor Schlüter [Allemagne] ; Wolfgang Engel [Allemagne]Source :
- Molecular Reproduction and Development [ 1040-452X ] ; 2001-04-01.
English descriptors
Abstract
During spermiogenesis, the successive replacement of somatic histones by basic proteins, the transition nuclear proteins and protamines, allows normal sperm condensation. Transition nuclear protein 2 (TNP2) is transcribed postmeiotically in round spermatids. In order to determine regulatory flanking sequences responsible for stage specific expression of TNP2 gene, different transgenic mice were generated by microinjection of fertilized eggs. We demonstrate here that 525 bp of 5′‐ and 920 bp of 3′‐flanking sequences of rat TNP2 gene could properly and efficiently direct chloramphenicol acetyltransferase gene expression to the postmeiotic male germ cells of transgenic mice. During male germ cell differentiation the first transgene transcripts were observed in round spermatids and translation started 6 days later in elongating spermatids, which is an evidence for posttranscriptional regulation of transgene expression. In contrast, transgenic mice bearing only the 525 bp 5′‐flanking sequences of rat transition protein 2 gene and 3′‐flanking sequences of the simian virus 40 large T antigen showed low levels of transgene expression in testis. From these results, it can be concluded that the 525 bp 5′‐flanking sequences regulate the cell specific expression and the sequences located in 920 bp 3′‐flanking region either enhance the transgene expression in the male germ cells or may have a posttranscriptional role in stabilizing the mRNA in addition to its function in delaying the mRNA translation. Using comparative alignment of 5′‐flanking of TNP2 genes from different species, the putative regulatory sequences are identified. Mol. Reprod. Dev. 58:368–375, 2001. © 2001 Wiley‐Liss, Inc.
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DOI: 10.1002/1098-2795(20010401)58:4<368::AID-MRD3>3.0.CO;2-J
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Dev.</title><idno type="ISSN">1040-452X</idno><idno type="eISSN">1098-2795</idno><imprint><publisher>John Wiley & Sons, Inc.</publisher><pubPlace>New York</pubPlace><date type="published" when="2001-04-01">2001-04-01</date><biblScope unit="volume">58</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="368">368</biblScope><biblScope unit="page" to="375">375</biblScope></imprint><idno type="ISSN">1040-452X</idno></series><idno type="istex">5EA824ACCB35120D40EBABF18E254E01A1273CA1</idno><idno type="DOI">10.1002/1098-2795(20010401)58:4<368::AID-MRD3>3.0.CO;2-J</idno><idno type="ArticleID">MRD3</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1040-452X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>haploid expression</term><term>transgenic mice</term><term>transition protein 2</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">During spermiogenesis, the successive replacement of somatic histones by basic proteins, the transition nuclear proteins and protamines, allows normal sperm condensation. Transition nuclear protein 2 (TNP2) is transcribed postmeiotically in round spermatids. In order to determine regulatory flanking sequences responsible for stage specific expression of TNP2 gene, different transgenic mice were generated by microinjection of fertilized eggs. We demonstrate here that 525 bp of 5′‐ and 920 bp of 3′‐flanking sequences of rat TNP2 gene could properly and efficiently direct chloramphenicol acetyltransferase gene expression to the postmeiotic male germ cells of transgenic mice. During male germ cell differentiation the first transgene transcripts were observed in round spermatids and translation started 6 days later in elongating spermatids, which is an evidence for posttranscriptional regulation of transgene expression. In contrast, transgenic mice bearing only the 525 bp 5′‐flanking sequences of rat transition protein 2 gene and 3′‐flanking sequences of the simian virus 40 large T antigen showed low levels of transgene expression in testis. From these results, it can be concluded that the 525 bp 5′‐flanking sequences regulate the cell specific expression and the sequences located in 920 bp 3′‐flanking region either enhance the transgene expression in the male germ cells or may have a posttranscriptional role in stabilizing the mRNA in addition to its function in delaying the mRNA translation. Using comparative alignment of 5′‐flanking of TNP2 genes from different species, the putative regulatory sequences are identified. Mol. Reprod. Dev. 58:368–375, 2001. © 2001 Wiley‐Liss, Inc.</div></front></TEI><affiliations><list><country><li>Allemagne</li></country><region><li>Basse-Saxe</li></region><settlement><li>Göttingen</li></settlement></list><tree><country name="Allemagne"><region name="Basse-Saxe"><name sortKey="Nayernia, Karim" sort="Nayernia, Karim" uniqKey="Nayernia K" first="Karim" last="Nayernia">Karim Nayernia</name></region><name sortKey="Bohm, Detlef" sort="Bohm, Detlef" uniqKey="Bohm D" first="Detlef" last="Böhm">Detlef Böhm</name><name sortKey="Engel, Wolfgang" sort="Engel, Wolfgang" uniqKey="Engel W" first="Wolfgang" last="Engel">Wolfgang Engel</name><name sortKey="Schluter, Gregor" sort="Schluter, Gregor" uniqKey="Schluter G" first="Gregor" last="Schlüter">Gregor Schlüter</name><name sortKey="Topaloglu, Ozlem" sort="Topaloglu, Ozlem" uniqKey="Topaloglu O" first="Özlem" last="Topaloglu">Özlem Topaloglu</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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