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Position: Home > Articles > Cloning and sequence analysis of muscovy duck reoviruses S14 and C4L8 strains NS gene Chinese Journal of Preventive Veterinary Medicine 2005,27 (2)

番鸭呼肠孤病毒非结构基因的克隆和序列分析

作  者：
张云;欧阳岁东;刘明;胡奇林;郎景华;童光志
单  位：
哈尔滨市兽医卫生防疫站黑龙江哈尔滨;中国农业科学院哈尔滨兽医研究所生物技术国家重点实验室;福建农科院牧医所
关键词：
番鸭呼肠孤病毒;NS基因;克隆;序列分析
摘  要：
参考GenBank禽呼肠孤病毒 (AvianReovirus,ARV)和番鸭呼肠孤病毒 (MuscovyDuckReovirus,MDRV)非结构基因 (NS)序列设计合成一对引物 ,对番鸭呼肠孤病毒S14和C4株NS基因进行RT_PCR扩增 ,克隆到pMD18_T载体中 ,并对克隆产物进行酶切鉴定和测序 ;番鸭呼肠孤病毒NS基因由 12 91bp核苷酸组成 ,与禽呼肠孤病毒NS基因相比 ,在非编码区第 115 5位少一个碱基 ,本文第一次证实 12 91bp是番鸭呼肠孤病毒NS基因特有的长度 ;番鸭呼肠孤病毒S14和C4株NS基因的 5’末端和 3’末端分别为 5‘GCTTTTT和TCATC_3’ ,是禽类呼肠孤病毒基因末端特有的碱基序列 ,S14和C4株NS基因的的有效阅读框 (2 4～ 112 7bp)编码 36 7个氨基酸组成的蛋白 ,分子量约为 4 0kDa ;番鸭呼肠孤病毒S14和C4株NS蛋白等电点分别是 7.3和 7.0 ,GC含量分别为 5 4 .2 6 %和 5 3.71% ,番鸭呼肠孤病毒S14和C4株NS基因间核苷酸同源性为 99.3% ,仅有 4个氨基酸差异 ,S14和C4与法国番鸭呼肠孤病毒 890 2 6株NS基因核苷酸同源性分别为 87.8%和 87.9% ,与鸡关节炎病毒S1133NS基因同源性分别为 79.0 %和 79.3% ;进化树分析表明本研究中的两株番鸭呼肠孤病毒非结构基因 (NS)与番鸭呼肠孤病毒的亲缘关系比禽呼肠孤病毒近的多 ,建议番鸭呼肠孤病毒应归属为正呼肠孤?
译  名：
Cloning and sequence analysis of muscovy duck reoviruses S14 and C4L8 strains NS gene
作  者：
ZHANG Yun~1, OUYANG Sui_dong~2, LIU Ming~1, HU Qi_lin~2, LANG Jing_hua~3, TONG Guang_zhi~1 (1.National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Harbin 50001,China;2.Fujian Husbandry and Veterinary Research Institute, Fujian 350003,China; 3.Harbin Veterinary Station,Harbin 150001,China)
关键词：
muscovy duck reovirus; NS gene; cloning; sequence analysis.
摘  要：
NS genes of muscovy duck reovirus S14 and C4 were amplified by RT_PCR with the primers designed by the reported avian and muscovy duck reoviruses. The PCR products were cloned to the pMD18_T vector and identified by enzyme cutting analysis. The sequence results showed that both NS genes are 1 191 bp. The NS conserved 5' and 3' terminal motifs of S14 and C4 are 5'_GCTTTTT and TCATC_3', which are the same as that of avian reoviruses. It contains one open reading frame that encodes a protein of 367 amino acids with a molecular mass of 40.0 kDa. Phylogenetic analysis shows that NS gene of S14,C4 and 89026 are all grouped with avian reoviruses. The homology of NS gene between S14 and C4 is 99.3 %. The NS genes of S14 and C4 are related to that of muscovy duck reoviruse 89026, respectively with 87.8 % and 87.9% homology. Compared to that of avian arthritis virus S1133,the homology of NS gene of S14 and C4 are respectively 79.0 % and 79.3 %. Generally, relationship of muscovy duck reovirus S14 and C4 to 89026 are more similar than that to avian arthritis virus S1133. The relationship of NS genes of muscovy duck S14 and C4 are more related to 89026 than to avian reovirus. The results of this study give the suggestion that the muscovy duck reoviruses might belong to one special gene group among avian reovirus of second subgroup of orthoreovirus genus.

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