Volume 2 Issue 6
Dec.  2006
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SHAO Yan-qing, ZHANG Dian-chang, SU Tian-feng, LU: Jun-lin, JIANG Shi-gui. The construction of pituitary gland cDNA library of Nibea coibor[J]. South China Fisheries Science, 2006, 2(6): 8-12.
Citation: SHAO Yan-qing, ZHANG Dian-chang, SU Tian-feng, LU: Jun-lin, JIANG Shi-gui. The construction of pituitary gland cDNA library of Nibea coibor[J]. South China Fisheries Science, 2006, 2(6): 8-12.
shu

The construction of pituitary gland cDNA library of Nibea coibor

  • 1.

    South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China

  • 2.

    Shanghai Fisheries University, Shanghai 200090, China

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  • Received Date: June 05, 2006
  • Revised Date: September 11, 2006
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    • Abstract

  • A cDNA expression library from the pituitary gland of Nibea coibor was constructed into pcDNA3.1 (+) eukarytical expression plasmid using SMARTTM cDNA library construction protocol. The anchor first strand cDNA containing asymmetrical SfiⅠ restriction enzyme sites (A & B) was synthesized by transcription of total RNA with the SMART technique. The long-distance(LD PCR) was performed using a modified oligo(dT) prime and anchor prime as the prime set, and anchor fist-strand cDNA as the template to enrich the cDNA population for full-length sequences. After digestion with SfiⅠ and size fractionation using CHROM SPIN-400 column, SMART cDNA was ligated into the SfiⅠ-digested pcDNA3.1 (+) -SfiⅠ vector and transferred into the competent cells of Escherichia coli TOP10 strain. The obtained N.coibor cDNA library contains 1.0×105 recombinants, the percentage of vectors with inserts was 90% and the average insert size was between 0.4~3.0 kb. The N.coibor cDNA library gives an ideal base for further study structure and characterization of the gene in relation to growth and development of N.coibor.

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    • References (9)
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