摘要
利用DD-RTPCR(正常鲤鱼外周血白细胞和经有丝分裂源刺激后白细胞的总RNA)获得的差异显示片段B10克隆(编码IL8的一段序列),对该基因进行DIG标记,对有丝分裂原刺激的鲤鱼外周血白细胞cDNA文库进行核酸杂交筛选,从0.8×104个重组噬菌体中,经过2轮筛选最终获得阳性克隆,挑选了3个分别命名为IL8-1,IL8-2,IL8-3。测序后经NCBI Blast分析表明,阳性克隆具有完整阅读框架,均为编码鲤鱼IL8的全长cDNA,编码的多肽由98个氨基酸残基组成,其相对分子质量理论推导值为10 848.9 Da,等电点为8.68。且与2003年12月Dev Comp Immunol.杂志报道的荷兰鲤鱼氨基酸的同源性达84%。
In order to study immune related genes in carp （ Cyprinus carpio L. ） leucocytes, cDNAs （ termed IL8 ） were cloned from the cDNA library of mitogen stimulated carp leucocytes by a probe gotten from the DD - RTPCR.. IL8 - 1, IL8 - 2 and IL8 - 3 cDNA respectively were 729bp, 753bp and 730bp in full length with an ORF of 294bp encoding IL8 gene. Those genes all encoded 98 amino acids with molecular weight of 10848. 9Da and isoeletric point of 8. 68. Blast searching revealed that there were significant homologues with interleukin-8 gene in carp leucocytes in Netherlands （reported in Dev Comp Immunol. ） amount to 84%.
出处
《云南农业大学学报》
CAS
CSCD
2008年第1期68-72,共5页
Journal of Yunnan Agricultural University
基金
国家自然科学资金资助项目(30200210)
关键词
鲤鱼
IL8
克隆
序列分析
Carp （Cyprinus carpio L. ）
interleukin-8 gene
Cloning
sequence analysis
