摘要
应用RT-PCR技术从ConA诱导培养的大熊猫外周血淋巴细胞总RNA中扩增得到大熊猫IL-4基因,并将其克隆到PGEM-T载体中。经菌落PCR鉴定、序列测定及序列分析,结果表明,克隆得到的IL-4基因开放阅读框由396个核苷酸组成,编码一个由132个氨基酸组成的多肽,包括编码24个氨基酸的信号肽和108个氨基酸的成熟肽。与GenBank中已登录的大熊猫IL-4（DQ166512）的核苷酸序列同源性分别为99.5%和100%;与其他哺乳动物如犬、猫、人、猪、牛、羊、马、兔、鼠等的IL-4核苷酸同源性为50.7%（鼠）～87.7%（犬）;IL-4编码氨基酸同源性在35.7%（鼠）～78.8%（犬）;进化树构建结果表明,大熊猫与犬、猫遗传距离最近。
By reverse transcription polymerrase chain reaction（RT-PCR）,giant panda IL-4 gene was cloned from total RNA which was extracted from ConA-stimulated-cultured peripheral blood lymphocytes of giant panda, the amplified products of PCR were cloned into PGEM-T vector. After identification and sequencing, the sequence analysis revealed that the open reading frame of giant panda IL-4 gene contained 396 nucleotides which encoding 132 amino acids,the signal peptide contained 24 amino acids and the mature IL-4 gene was composed of 108 amino acids. Compared with one published sequence of giant panda（DQ166512） ,the homology of nucleotide sequence and deduced amino acid sequence were 99.5%and 100% respectively; compared with other mammals in GenBank,including dog, cat,human,pig,cow, horse,goat, rabbit, rat and so on, the homology of nucleotide sequence was between 50. 7% （rat）- 87.7 % （dog）, the homology of amino acid sequence was between 35.7 % （rat） - 78. 8 % （dog）. Phylogenetic analysis based on the nucleotide sequence of the IL-4 gene showed that giant panda had the most closed genetic distance with dog and cat.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2008年第5期549-552,共4页
Chinese Journal of Veterinary Science
基金
成都大熊猫繁育研究基金会资助项目(CPF008)
