中国人4型登革病毒E基因部分片段的cDNA克隆与序列分析  

The cloning and sequence analysis of cDNA of the partial E genomic region of Dengue type 4 virus from a Chinese patient

作　　者：李刚[1] 姚集鲁[1] 彭文伟[1] 王飞[1] 

机构地区：[1]中山医科大学传染病学教研室

出　　处：《中华实验和临床病毒学杂志》1997年第1期62-65,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：广东省委基金

摘　　要：１９９３年１０月广东省发生登革热的流行，我们从其中一个病人血清中扩增到４型登革病毒（ＤＶ４）包膜蛋白基因（Ｅ）的部分ｃＤＮＡ片段，对其进行了分子克隆及序列测定。ＤＶＲＮＡ经随机引物逆转录后用ＤＶ４特异的引物扩增，获得４２１ｂｐ的产物，补齐和提纯后插入ｐＵＣ１８和ｐＵＣ１９质粒载体，采用双脱氧链末端终止法测定核苷酸序列，与同型ＤＶ代表株比较，同源性达到９３７２％，与不同型ＤＶ的同源性介乎６１２６％～６４４０％，而与同属的乙型脑炎病毒比较，同源性只有４０３１％。比较推导的氨基酸序列后发现一个含有１２个氨基酸的保守区，可能是黄病毒属中具有重要生物学功能的部位。The partial envelope (E) protein gene of Dengue virus type 4 (DV4), derived from the serum of a patient with dengue fever in an epidemic during October 1993 in Guangdong Province of China, was cloned and sequenced. DV RNA was converted to cDNA by reverse transcription with random primers before the polymerase chain reaction using DV4 specific primers was performed. The amplified product of 421bp was subsequently filled in 3' recessed ends, isolated, purified and inserted into pUC18 and pUC19 plasmid vector. Their sequences were determined by dideoxy nucleotide chain termination method. A comparison between DV4 Chinese strain and several other previously reported strains shows the homology to be 93.72% for the same dengue serotype, 61.26～64.40% for the other dengue serotypes, only 40.31% for Japanese encephalitis virus, which belongs to the member of flavivirus family. A conserved sequence with 12 amino acids observed from deduced protein sequence probably represents an essential functional element among flavivirus family.

关 键 词：登革热病毒 克隆 分子 碱基序列 CDNA 

分 类 号：R373.33[医药卫生—病原生物学]