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Journal of plant biotechnology v.7 no.1, 2005년, pp.51 - 55  

Unusual Orientation of cDNAs Found in a cDNA Library

Lee Jeongyeo    (Genome Research Institute, Chungnam National University   ); Song Hayoung    (Genome Research Institute, Chungnam National University   ); Lim Yong-Pyo    (Genome Research Institute, Chungnam National University   ); Hur Yoonkang    (Genome Research Institute, Chungnam National University  );
  • 초록

    Many cloning vectors in which cDNAs can be inserted to the sense orientation have been developed. Uni-ZAP XR vector (Stratagene) should contain clones that are oriented to sense direction with respect to T3 RNA polymerase primer. Unexpectedly, large portions of cDNAs in Chinese cabbage cDNA library showed unusual insertions, antisense orientation and a hybrid of two different clones. Using two clones, 4H03 and 53-B10, derived from different cDNA libraries, we proposed and demonstrated the possibility of unusual-construct formation by in vitro translation and northern blot analysis. The 4H03 clone was inserted with inverse direction, and its transcript and translation product could be produced by T7 RNA polymerase, indicating that this clone is definitely inserted into inverse orientation. The 53-B10 that contains two independent genes was turned out to be a hybrid in which two genes are inserted to opposite direction each other. All unusual constructs might be due to the presence of small fragments of DNA, like adapter. However, the mechanism underlined the formation of unusual constructs is still remain to be solved.


  • 주제어

    directional cloning .   EeoRI adapter .   cDNA library .   Chinese cabbage.  

  • 참고문헌 (11)

    1. Baltimore D (1970) RNA-dependent DNA polymerase in virions of RNA tumour viruses. Nature 226: 1209-1211 
    2. Rougeon F, Kourilsky P, Mach B (1975) Insertion of a rabbit ${\beta}$ -globin gene sequence into an E. coli plasmid. Nucleic Acids Res 2: 2365-2378 
    3. Ross J, Aviv H, Scolnick E, Leder P (1972) In vitro synthesis of DNA complementary to purified rabbit globin mRNA (RNA-dependent DNA polymerase-reticulocyte-hemoglobindensity grdient centrifugation-oligo (dT) primer). Proc Natl Acad Sci 69: 264-268 
    4. Krug MS, Berger SL (1987) First-strand cDNA synthesis primed with oligo (dT). Method Enzymol 152: 316-325 
    5. Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Press, Cold Spring Harbor, New York 
    6. Rabbius TH (1976) Bacterial cloning of plasm ids carrying copies of rabbit globin messenger RNA. Nature 260: 221-225 
    7. Kretz PL, Reid CH, Greener A, Short JM (1989) Effect of lambda packaging extract mer restriction activity on DNA cloning. Nucleic Acids Res 17: 5409 
    8. Temin HM, Mizutani S (1970) RNA-dependent DNA polymerase in virions of Rous sarcoma virus. Nature 226: 1211-1213 
    9. Verma IM, Temple GF, Fan H, Baltimore D (1972) In vitro synthesis of DNA complementary to rabbit reticulocyte 10S RNA. Nat New Biol 235: 163-167 
    10. Bank A, Terada M, Metafora S, Dow L, Marks PA (1972) In vitro synthesis of DNA components of human genes for globins. Nat New Biol 235: 167-169 
    11. Short JM, Fernandez JM, Sorge JA, Huse ED (1988) ${\lambda}$ ZAP: a bacteriophage ${\lambda}$expression vector with in vivo excision properties. Nucleic Acids Res 16: 7583-7600 

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