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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 25, No. 2, 2004, pp. 166-171
Bioline Code: zr04030
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 25, No. 2, 2004, pp. 166-171

 en Construction of a Skin cDNA Library of Cashmere Goat and Cloning of KAP6-2 Full-length cDNA
YIN Jun HU Tin-mao LI Jin-quan ZHANG Chun-lan GUO Zhi-zheng ZHOU Huan-min

Abstract

In order to understand the molecular mechanism of the characteristic of cashmere,we constructed a cDNA library using the SMART cDNA library construction kit (Clontech).Total RNA was isolated from the goat ( Capra hircus check for this species in other resources ) skin tissue with hair follicle angen.Oligotex (QIAGEN) was used to isolate mRNA from total RNA.The "anchor first-strand cDNA" synthesized by reverse transcription with the SMART technique.The LD-PCR was performed using a modified oligo (dT) primer and an anchor primer as the primer set,and anchor first-strand cDNA as the template to enrich the cDNA population for full-length sequences.After digestion with Sfi I and size fractionation,SMART cDNA was ligated into the Sfi I-digested pBluescript II SK (with Sfi I A and B site).The ligation mixture was transformed into E.Coli 5α.The cDNA library contained 1.8x105 independent clones.Randomly select cDNA clones and sequence with the 5 prime end,a full-length KAP6-2 cDNA was found by compared with those in the NCBI database (nr) using the Blast-N programs,it showed 75.5% identity in amino acids with mouse KAP6-2 and the accession number in GenBank is AY316158.

Keywords
Cashmere goat; KAP6-2; Full-length cDNA

 
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