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应用3’RACE技术扩增仿刺参C型凝集素基因及实验条件的优化 被引量:2
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作者 李莹 韩璐璐 《辽宁师范大学学报(自然科学版)》 CAS 2012年第2期246-251,共6页
从GenBank上调取刺参C型凝集素(C-type Lectin)基因序列EST,根据此序列设计RACE引物,采用PCR扩增技术得到了仿刺参C-type Lectin基因序列(EST),根据这段EST序列设计1个基因特异引物(GSPF),与通用引物(UPM)扩增,成功地克隆到了该基因的3... 从GenBank上调取刺参C型凝集素(C-type Lectin)基因序列EST,根据此序列设计RACE引物,采用PCR扩增技术得到了仿刺参C-type Lectin基因序列(EST),根据这段EST序列设计1个基因特异引物(GSPF),与通用引物(UPM)扩增,成功地克隆到了该基因的3’末端序列.同时,对仿刺参C型凝集素基因3’克隆的实验条件进行了优化.该扩增片段长度为670bp,与已知序列重叠部分为417bp.经测序和比对发现该段序列与预期的目标基因的序列一致. 展开更多
关键词 仿刺参 C型凝集素 3’race技术
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Cloning and Expression of a Profilin Gene from Rapeseed 被引量:4
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作者 叶秋 李旭锋 +3 位作者 徐莺 王劲 林娟 陈放 《Acta Botanica Sinica》 CSCD 2001年第7期727-730,共4页
Profilin has recently been identified as an actin-binding protein in higher plants. A cDNA clone (designated Repro) encoding profilin gene was isolated from rapeseed ( Brassica napus L. cv. canadian Tween) using RT-PC... Profilin has recently been identified as an actin-binding protein in higher plants. A cDNA clone (designated Repro) encoding profilin gene was isolated from rapeseed ( Brassica napus L. cv. canadian Tween) using RT-PCR technique. Sequence analysis showed 82% similarity to Zea mays L. ZmPro3, 85% to Arabidopsis AthPRF1, 82% to Nicotiana tabacum L. NTPRO, 81% to Oryza sativa L. profilin A. A new full-length cDNA was obtained by 5'-RACE and 3'-RACE techniques. Sequence analysis showed that the size of full-length cDNA is 672 bp which contains a major open reading frame of 134 amino, acids, 5' and 3' untranslated regions and a long Poly (A) tail. Northern blot analysis showed that the profilin gene is a pollen and anther specific gene. 展开更多
关键词 profilin gene 3 '-race 5 '-race rapeseed pollen RT-PCR
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