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驯鹿β-防御素-1 cDNA 3′末端的克隆及序列分析
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郝永峰1，杨银凤1，赵艳芳2，余兴帮3，都格尔斯仁1
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（1.内蒙古农业大学 动物胚胎与发育生物学研究室，内蒙古 呼和浩特010018； 2.呼伦贝尔市兽医站，
内蒙古 海拉尔012008； 3.呼伦贝尔市鄂温克旗农牧业局，内蒙古 鄂温克旗012000）
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摘要：
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为进一步研究驯鹿β-防御素-1(reBD-1)基因的分子结构，利用已克隆出的reBD-1部分片段，设计了1条特异性上游引物，并以反转录引物中的部分序列即3 sites Adaptor Primer作为下游引物，采用3′RACE技术成功克隆了reBD-1 cDNA的3′末端序列。通过与已知reBD-1片段拼接，得到了192bp的完整开放读码框（ORF）、终止密码子TAA、118bp的3′非翻译区（3′UTR）以及poly（A）15，其中ORF编码具有64个氨基酸残基的reBD-1前原肽。 |
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关键词：
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β-防御素； 3′RACE； 驯鹿； 克隆 |
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中图分类号：S 852.42:Q 523.8 文献标识码：
A 文章编号：1673-4696(2006)03-0239-04 |
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Molecular cloning and analysis of 3′end sequence of
reindeer β-defensin-1 cDNA by 3′RACE |
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HAO Yong-feng1 , YANG
Yin-feng1, ZHAO Yan-fang2
,YU Xing-bang3 ,Dugeersiren1 |
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(1.Laboratory of Animal Embryology and developmental biology, Nei Monggol Agricultural University,
Hohhot 010018, China; 2. Hulun-Beier Prefectural Station of Veterinary Medicine, Hailaer, 012008, China；
3. Ewenke County Bureau of Agriculture and Animal Husbandry, Hulun-Beier, Ewenke 012000, China) |
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Abstract: In order to study the molecular structure of reindeer β-defensin-1(reBD-1) gene, the authentic 3′end sequence of reindeer β-defensin-1(reBD-1) gene was successfully cloned by the 3′RACE technique with a specific upper primer designed according to the known
reBD-1 partial cDNA fragment and the 3 sites Adaptor Primer as reverse primer. By joining the 3′end sequence with the known
reBD-1 partial sequence, the reBD-1 gene was cloned which contained an ORF of 192bp, a stop codon of TAA, a 3′ untranslated region (3′UTR) of 118bp and poly(A)15. The ORF of 192bp encoded a
64-amino acid prepro-peptide. |
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Key words: β-defensin; 3′RACE; reindeer; cloning |
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