摘要
实验应用反转录-聚合酶链式反应(RT-PCR)技术,在国内首次从ConA诱导培养的中国大耳白肉兔外周血淋巴细胞总RNA中扩增得到IFN-γ、IL-2和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:①经克隆得到的IFN-γ基因(序列号:DQ852341),与Genbank中已登录的欧洲兔IFN-γ基因(序列号:AB010386)的核苷酸序列和推知氨基酸序列的同源性均为100%;与其他哺乳动物如犬、猫、人、猪、牛、羊、马、大熊猫和鼠等的核苷酸同源性在63.0%(鼠)～77.6%(人)之间;编码氨基酸同源性在41.7%(鼠)～65.7%(人)之间;②扩增得到的IL-2基因(序列号:DQ852342),与Genbank中已登录的欧洲兔IL-2基因(序列号:AF068057)的核苷酸序列和推知氨基酸序列的同源性分别为99.6%和100%;与其他哺乳动物如犬、猫、人、猪、牛、羊、马、大熊猫和鼠等的核苷酸同源性在64.8%(鼠)～86.0%(人)之间;编码氨基酸同源性在55.7%(鼠)～80.1%(人)之间;③扩增得到的IL-4基因(序列号:DQ852343),与Genbank中已登录的欧洲兔IL-4基因(序列号:AF169169)的核苷酸序列和推知氨基酸序列同源性均为100%;与其他哺乳动物如犬、猫、人、猪、牛、羊、马、大熊猫和鼠等的核苷酸同源性在57.0%(大熊猫)～69.8%(人)之间;编码氨基酸同源性在43.0%(鼠)～53.6%(人)之间。用这3个基因分别构建的进化树结果都表明,兔与人的亲缘关系相对较近,与鼠的亲缘关系最远,这与传统的分类地位基本吻合,即兔与鼠应分别归为兔形目和啮齿目动物。
With reverse transcription polymerase chain reaction （RT- PCR）, IFN- γ, IL - 2 and IL -4 gene were gained from total RNA which was extracted from ConA-stimulated-cultured peripheral blood lymphocytes of Chinese white rabbit. The sequence analysis revealed that： ①Compared with one published sequence （sequence number： AB010386）, the homology of nucleotide sequence and amino acid sequence of IFN-γgene （sequence number： DQ852341） were both 100 % ; compared with other mammals in Genbank, including dog, cat, human, pig, cow, horse, goat, giant panda, rat, and so on, the homology of nucleotide sequence was between 63.0% （Rat） and 77.6% （Human）, the homology of amino acid sequence was between 41.7% （Rat） and 65.7% （Human） ; ②Compared with one published sequence （sequence number： AF068057）, the homology of nucleotide sequence and amino acid sequence of IL - 2 gene （sequence number： DQ852342） were 99.6 % and 100 % respectively; compared with other mammals in Genbank, including dog, cat, human, pig, cow, horse, goat, giant panda, rat, and so on, the homology of nucleotide sequence was between 64.8% （Rat） and 86.0% （Human）, the homology of amino acid sequence was between 55.7% （Rat） and 80.1% （Human） ; ③Compared with one published sequence （sequence number： AF169169）, the homology of nucleotide sequence and amino acid sequence of IL - 4 gene （sequence number： DQ852343） were both 100%; compared with other mammals in Genbank, including dog, cat, human, pig, cow, horse, goat, giant panda, rat, and so on, the homology of nucleotide sequence was between 57.0% （Giant panda） and 69.8% （Human）, the homology of amino acid sequence was between 43.0% （Rat） and 53.6% （Human）. Phylogenetic analysis based on the nucleotide sequences of the IFN-7, IL-2 and IL-4 gene respectively showed that rabbit had more closed genetic relation with human, but the farthest with rat. It was consistent with the orthodox taxonomic results, rabbit and rat were belonged to Lagomorpha and Rodentia respectively.
出处
《四川农业大学学报》
CSCD
2007年第4期468-474,共7页
Journal of Sichuan Agricultural University
基金
四川省学术带头人培养基金项目
作者简介
通讯作者（Corresponding author）。
