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PCR-generated cDNA libraries from reduced numbers of mouse oocytes

Published online by Cambridge University Press:  26 September 2008

Frédérique Revel ,
Jean-Paul Renard  and
Véronique Duranthon
Frédérique Revel*
Affiliation:
Unité de biologie du développement INRA, Jouy en Josas, France
Jean-Paul Renard
Affiliation:
Unité de biologie du développement INRA, Jouy en Josas, France
Véronique Duranthon
Affiliation:
Unité de biologie du développement INRA, Jouy en Josas, France
*
F. Revel, Unité de biologie du développement, INRA, F78352 Jouy en Josas, France. Telephone: (33) 1 34652592. (33) 1 34652877.0.
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Summary

We describe a rapid and reproducible method for cloning cDNA amplified from 10 mouse oocytes. The procedure consists in priming cDNA synthesis from a crude cellular extract using an oligo d(T) containing primer and submitting the size-limited cDNA first strand to poly(dG) tailing. The whole cDNA population is then polymerase chain reaction (PCR) amplified using two primers complementary to oligo d(A) and oligo d(G) ends of the cDNA. In this procedure no purification steps are required. We obtained about 5 ×106 clones from 10 oocytes. Screening of the library showed that the relative abundance of the transcripts was preserved during amplification and cloning and that the procedure allows cloning of low-abundance sequences at least as rare as 0.008% of the mRNA. The repeatable generation of representative cDNA libraries from reduced numbers of oocytes or embryos should open new opportunities for obtaining genetic information from mammalian preimplantation embryos.

Keywords

DNA libraryMammalian oocytespolymerase chain reactionPreimplantation embryos
Type
Article
Information
Zygote , Volume 3 , Issue 3 , August 1995 , pp. 241 - 250
Copyright
Copyright © Cambridge University Press 1995

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References

Bachvarova, R. (1988). Small B2 RNAs in mouse oocytes, embryos, and somatic tissues. Dev. Biol. 130 513–523.CrossRefGoogle ScholarPubMed
Bachvarova, R., De Leon, V., Johnson, A., Kaplan, G. & Paynton, B.V. (1985). Changes in total RNA, polyadenylated RNA, and actin mRNA during meiotic maturation of mouse oocytes. Dev. Biol. 108, 325–331.CrossRefGoogle ScholarPubMed
Beliavsky, A., Vinogradova, T. & Rajewsky, K. (1989). PCR based cDNA library construction: general cDNA libraries at the level of a few cells. Nucleic Acids Res. 17, 2919–32.CrossRefGoogle Scholar
Brady, G.Barbara, M.& Iscove, N. (1990). Representative in vitro cDNA amplification from individual hematopoietic cells and colonies. Methods Mol. Cell. Biol. 2, 17–25.Google Scholar
Brunet, J.F.Shapiro, E.Foster, S.A.Kandel, E.R.& Iino, Y. (1991). Identification of a peptide specific for Aplysia sensory neurons by PCR-based differential screening. Science. 252, 856–9.CrossRefGoogle ScholarPubMed
Church, G.M.& Gilbert, W. (1984). Genomic sequencing.Proc. Natl. Acad. Sci. USA. 81, 1991–5.CrossRefGoogle ScholarPubMed
Clegg, K.B.& Piko, L. (1983). Quantitative aspects of RNA synthesis and polyadenylation in 1-cell and 2-cell mouse embryos. J. Embryol. Exp. Morphol. 74, 169–82.Google ScholarPubMed
Copp, A. (1995). Death before birth: clues from gene knockouts and mutations. Trends Genet. 11, 87–93.CrossRefGoogle ScholarPubMed
Dumas, B. (1980). Construction de plasmides en vue de l'étude de la notion de promoteur chez les eucaryotes. Diplome d'éetude approfondie de biochimie fondamentale, University of Paris VII.Google Scholar
Feinberg, A.P.& Vogelstein, B. (1984). A technique for radio-labeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137, 266–7.Google Scholar
Fregien, N.Dolecki, G.J.Mandel, M.& Humphreys, T. (1993). Molecular cloning of live individual stage and tissue-specific mRNA sequences from sea urchin pluteus embryos. Mot. Cell. Biol. 3, 1021–31.Google Scholar
Goldman, D.S., Kiessling, A.A., & Cooper, G.M. (1988). Post-transcriptional processing suggest that c-mos functions as a maternal message in mouse eggs. Oncogene 3, 159–62.Google ScholarPubMed
Keshet, E.Rosenberg, M.P.Mercer, J.A.Propst, F.Vande Woude, G.F.Jenkins, N.A.& Copeland, N.G. (1988). Developmental regulation of ovarian-specific mos expression. Oncogene 2, 235–40.Google ScholarPubMed
Lalanne, J.L.Bregegere, F.Delarbre, C.Abaatado, J.P.Gachelin, C.& Kourilsky, p. (1982). Comparison of nucleotide sequences of mRNAs belonging to the mouse H-2 multigene family. Nucleic Acid Res. 10, 1039–48.CrossRefGoogle Scholar
Latham, K.E.Garrels, J.L.Chang, C.& Solter, D. (1991). Quantitative analysis of protein synthesis in mouse embryos. I. Extensive reprogramming of the one- and two-cell stages. Development 112, 921–32.CrossRefGoogle ScholarPubMed
Latham, K.E.Garel, J.I.& Solter, D. (1994). Alterations in protein synthesis following transplantation of mouse 8-cell stage nuclei to enucleated 1-cell embryos. Dev. Biol. 163, 341–50.CrossRefGoogle ScholarPubMed
Luqmani, Y.A.& Lymboura, M. (1994).Subtraction hybridization cloning of RNA amplified from different cell population microdissected from cryostat tissue section. Anal. Biochem. 222, 102–9.CrossRefGoogle Scholar
Maniatis, T.Fritsch, E.F.& Sambrook, J. (1982). Molecular Cloning: A laboratory Manual. New York: Cold Spring Harbor Laboratory.Google Scholar
McConnel, J.& Watson, C.J. (1986). Construction of a representative cDNA library from mRNA isolated from mouse oocytes. FEBS Lett. 195, 99–203.CrossRefGoogle Scholar
Miyamoto, H.Matsushiro, A.& Nozaki, M. (1993). Molecular cloning of a novel mRNA sequence expressed in cleavage stage mouse embryos. Mol. Reprod. Dev. 34 1–7.CrossRefGoogle ScholarPubMed
Moreau, J.Touzalen, N. & Mechali, M. (1995). Isolation of cDNAs from maternal mRNAs specifically present during early development. Mol. Reprod. Dev. (in press).CrossRefGoogle ScholarPubMed
Nozaki, M.Murata, K.Morita, T.& Matsushiro, A. (1988). Isolation of ENDO A cDNA from mouse 8-cell stage embryos. Biochem. Biophys. Res. Commun. 154, 890–4.CrossRefGoogle Scholar
Paris, J.& Philippe, M. (1990). Poly(A) metabolism and polysomal recruitment of maternal mRNAs during early Xenopus development. Dev. Biol. 140, 221–4CrossRefGoogle ScholarPubMed
Piko, L.& Clegg, K.B. (1982). Quantitative changes in total RNA, total poly(A) and ribosomes in early mouse embryos. Dev. Biol. 89, 362–78.CrossRefGoogle ScholarPubMed
Poirier, F.Chan, C.T.J.Timmons, P.M.Robertson, E.J.Evans, M.J.& Rigby, P.W. (1991). The murine H19 gene is activated during embryonic stem cell differentiation in vitro and at the time of implantation in the developing embryo. Development. 113, 1105–4.CrossRefGoogle ScholarPubMed
Richoux-Duranthon, V. (1991). Rôle des effets parentaux dana le developpement de l'embryon. Etude à partir d'un système modèle: Is lignèe de souris DDK. Thesis, University of Paris VI.Google Scholar
Rothstein, J.L.Johnson, D.Deloia, J.A.Skowronski, J.& Solter, D. (1992). Gene expression during preimplantation mouse development. Genes Dev. 6, 1190–201.CrossRefGoogle ScholarPubMed
Sargent, T.D.& Dawid, I.G. (1983). Differential gene expression in the gastrula of Xenopus laevis. Science 222, 135–9.CrossRefGoogle ScholarPubMed
Smith, D.& Gridley, T. (1992). Differential screening of a PCR-generated mouse embryo cDNA library: glucose transporters are differentially expressed in early postimplantation mouse embryos. Development. 161, 555–61.CrossRefGoogle Scholar
Swalla, B.J.Makabe, K.W.Satoh, N.& Jeffery, R.W. (1993). Novel genes expressed differentially in ascidians with alternate modes of development. Development. 119, 307–18.CrossRefGoogle ScholarPubMed
Taylor, K.D.& Piko, L. (1987). Pattern of mRNA prevalence and expression of B1 and B2 transcripts in early mouse embryos. Development. 101, 877–92.CrossRefGoogle ScholarPubMed
Temeles, G.L.Ram, P.T.Rothstein, J.L.& Schultz, R.M. (1994). Expression patterns of novel genes during mouse preimplantation embryogenesis. Mol. Reprod. Dev. 37, 121–9.CrossRefGoogle ScholarPubMed
Thompson, E.M.Christians, E.Stinnakre, M.G.& Renard, J. P. (1994). Scaffold-attachment regions stimulate HSP70.1 expression in mouse preimplantation embryos but not in differentiated tissues. Mol. Cell. Biol. 14, 4694–703.Google ScholarPubMed
Urven, L.E.Weng, D.E.Schumaker, A.L.Gearhart, J.D.& McCarrey, J.R. (1993). Differential gene expression in fetal mouse germ cells. Biol. Reprod. 48, 564–74.CrossRefGoogle ScholarPubMed
Varmuza, S.& Tate, p. (1992). Isolation of epiblast-specific cDNA clones by differential hybridization with polymerase chain reaction-amplified probes derived from single embryos. Mol. Reprod. Dev. 32, 339–48.CrossRefGoogle ScholarPubMed
Welsh, J.Liu, J.P.& Efstratiadis, A. (1990). Cloning of PCR-amplified total cDNA: construction of a mouse oocyte cDNA library. Genet. Anal. Techn. Appl. 7, 5–17.CrossRefGoogle ScholarPubMed
Weng, D.E.Morgan, R.A.& Gearhart, J.D. (1989). Estimates of mRNA abundance in the mouse blastocyst based on cDNA library analysis. Mol. Reprod. Dev. 1, 233–41.CrossRefGoogle ScholarPubMed