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1992 Vol.07(3)



Advances in Molecular Biology of Infectious Bursal Disease Virus

CHEN Shi-You, ZHANG Zi-Jun

1992, 7(3): 241

The Viral Inclusion Bodies in the Autopsy Tissues of Patients with Epidemic Hemorrhagic Fever

YANG Shou-Jing, LIU Yan-Fang, HAN Yan-Ping, XU Zhi-Kai, YAN Pei-Song

1992, 7(3): 246

Using polyclonal antibody (PAb)and monoclonal antibodies(MAbs) against epidemic hemorrhagic fever virus (EHFV), the viral antigens in fresh and long-term storage EHF autopsy tissues were studied comparatively with more sensitive multi-layer PAP and PAP combined with ABC (PAP+ABC) techniques. The results showed that the inclusion bodies (IB) with viral antigens positive could be demonstrated in the autopsy tissues. Four types of IB, designated solid. small-bubble, big-bubble and multi-bubble, could be observ...

Clone and Expression of Human Papillomavirus Type 16 L_1 Open Reading Frame

XU Xiu-Ping, ZHAO Wei-Meng, DONG Jie-De, ZHAO Li-Xin, CHEN Chen-Hua, StevenA.Jenison

1992, 7(3): 253

人乳头瘤病毒16型(HPV16)含有两个晚期开放读码框(L_1ORF和L_2ORF),L_1ORF编码主要衣壳蛋白,我们用质粒pHPV16、p16L_2BX_5和pATH,采用基因重组技术,制备了含有HPV16个长L_1ORF序列的基因克隆p16L_1BN(5071—253),它能在大肠杆菌中有效表达,产生分子量约90KD的含有E. coli trpE的融合蛋白。Western blot检测,该蛋白可被抗牛乳头瘤病毒的抗体识别,这说明克隆p16L_1BN能有效表达,基因产物具有乳头瘤病毒型的共同抗原的性质。

Detection of Human Papillomavirus Type 16 DNA in Head and Neck Tumors

YANG Ping, DIAO Wen-Xian, LIU Xue-Feng, WU Xin-Xing, DING Xiao-Hua, YAO Chang-De, YANG Qiang

1992, 7(3): 259

In order to probe the relatignship between human papillonavirus type 16 and head-neck tumors, using fresh tissue and paraffin-embeded tissue sections preparing DNA to detect HPV16 DNA in head and neck tumor, by PCR and hybridization technique. The results showed that HPV16 DNA sequences were positively amplified in one laryngeal papilloma (11.1%), in 3 inverted papillomas of the nose (42.9%). in 4 laryngeal squamous cell earcinomas(20%).in 3 oral eaneers(27.3%). LSCC of glottie origin showed higher PHV16 DN...

Mutagenesis and Cloning of HIV-1 gag Gene Fragment by Polymerase Chain Reaction

JI Chang-Hua, SU Cheng-Zhi, YAN Xiao-Jun

1992, 7(3): 265

In the present paper. the primers PG01 and PG02 were synthesized in which the EcoRI and BamHI recognition sites as well as the initiation and termination codons were designed for convenience of subsequent cloning and expression. Micrograms of target gene sequence was produced by 30 rounds of amplification with pJG 423 as a template. The amplified gene fragment was digested with both EeoRI and BamHI and ligated to pUC19 predigested with the same restriction endonucleases. The recombinant plasmid pCY52 was id...

Observation on the Persistance of Immunity in Humans Vaccinated with Hepatitis B Vaccine

ZHAO Wen-Bin, ZHOU Ke-Jie, WU Tao-Lin

1992, 7(3): 272

The immune persistance of three batches of domestic Hepatitis B Vaccine at a dose of 10μg were studied for four years. The results showed that in the same age group there was no difference in immune persistance by sex. The critical protective rate and effective protective rate were higher in the infant groups than in the adult groups. The difference has statistical significance (P<0.05). The protective efficacy of primary vaccination in the infants group dropped to lower than 60 per cent after four years...

Immunity Studies on the Euproctis pseudoconspersa Nuclear Polyhedrosis Virus

DAN Zheng-Li, ZHANG Li-Ren, CHEN Dai-Hua

1992, 7(3): 276

Methods such as immunodiffusion. counter-immunoelectrophoresis. enzyme-linked immunosorbent assay (ELISA) and solidphase immune electron microscopic(SPIEM) technique were used to study the antigenicity of EpNPV and relationships between EpNPV and other ten kinds of NPV Results indicated that the antiserum of EpNPV reacted only with the purified particles of EpNPV, and did not produce any immunoreaction with polyhedron protein and other ten kinds of NPV. Such tests suggested that there may be no serological ...

Effect of the Nuclear Polyhedrosis Virus from Tea Geometrid (Ectropis obliqua hypulina) on the Reproduction of tea Geometrid under High Temperature

YE Gong-Yin, HU Cui, SHU Dun-Qing, SHANG Jian-Nong

1992, 7(3): 283

Effect of the nuclear polyhedrosis virus from Ectropis oblioua hypulina Wehrli (EoNPV) on the reproduction of tea geometrid under high temperature condition in July and August was reported. The results indicated: The development rate of ovaries in female pupae or adults survived from the viral infection was no significantly different from that of the healthy ones; Fed with the virus at the late of the 3rd instar, the beginning of the 4th or 5th instar, both of the pupation and emergence rate were reduced di...

Study on the Infective Characters of Buzura suppressaria Nuclear Polyhedrosis Virus in BS484 Cell Line

XU Xu-Shi, XIE Tian-En

1992, 7(3): 289

This thesis reported the following infective charaeters of Buzura suppressatia nuclear polyhedrosis virus (BsNPV) in Bs484 cell line: 1. The viral infection rate was the highest when the virus was inoculated in Bs484 cells had been subeultured for 3—4 days. 2. The viral inocuiating quantities paralleled the total yield of polyhadra in virus-infected cells in certain range. 3. The viral tite had (?)o significant change after BsNPV was serially passaged for sevan times in Bs484 cells. 4. The synthesis of the ...

Cytopathic Changes of Host Infected by Euproctis pseudoconspersa Nuclear Polyhedrosis Virus

JU Tong, ZHANG Li-Ren

1992, 7(3): 296

Cytopathology of mjdgut cells of Euproctis pseudoconspersa infected with a nuclear polyhedrosis virus was studied by transmission electron microscope, seannig electron microscope DMSO cracking method, ultramicrotome, energy dispersive X-ray microanalysis and negative staining method. Between 48—120 hours infected with EpNPV, host cellular organs had apparently pathological changes. Nucleus enlarged, virogenie stroma existed within the nucleus. The number of mitoehondria increased, mitochondria hypertrphym i...

New Technique for the Production of Viral Insecticide of B. suppressaria Nuclear Polyhedrosis Virus

XIE Tian-En, BANG Hui-Yin, CENG Yun-Tian, ZHANG Zhong-Shen, WANG Lu-Meng, ZHANG Yang-Lian, LIU Yu-Lan, JIN Feng, LIU Song-Hua, DENG Wan-Gui, XIAO Zuo-Lv

1992, 7(3): 304

The oil tree moth (B. suppressaria larvae) was grown to 4 instar using a seml-synthetic artificial diet. then infected with (BsNPV) inoculum propagated in the insect cell cultures. 7 days p.i., the virus-infected larvae were collected and virus (PIB) was purified by differential centrifugation. Then, a flowable formulation was made with the glycerin, virus-protective agent, emulsion and antibiotic preparation was determined by standard bioassays using laboratory-reared larvae on artificial diet. LC50 was 2....

Kinds of Virus Causing Viral Disease on Sichuan Zha-Cai (Brassica juncaa var. tumida) as Detected by ELISA Method

LI Xin-Yu, CAI Yue-Song, WANG Bin, YU Jia-Lan

1992, 7(3): 311

In 1990—1991. from the fields in near- and-far suburb of nine counties or municipalities in Siehuan province, 492 accessions of Brassica juncea plant showing viral disease symptom were collected both in the initial and the prevalent period of the disease and were subjected to indirect ELISA tests with 6 kinds of antiserum—— TuMV, CMV. TMV. PVX. PVY. CaMV—— in an attempt to find their causal agent. 422 speeimens, accounting for85.77%, of the total, showed reaction with TuMV antiserum or with antiserum of one...

Screening and Application of Differential Hosts to Cucumber Mosaic Virus Strains on Pepper

YANG Yong-Lin, YAN Su-Zhen, WANG Hui, CHEN Zheng-Hua

1992, 7(3): 317

According to Flor s "Gene to Gene" theory and Omaro and Tomaru s method to distinguish CMV strains, we studied "strains of gene type" and "strains of pathogenetic type" 5 varities were selected as differential variety spectrum from 373 sweet or hot pepper, They were numbered LS-8501 (HR), LS-8502(R), LS-8503(T), LS-8504(S) and LS-8505(HS). 59 CMV isolates were divided into 5 strains by the spectrum, They were divided into 5 strains by the spectrum and named CMV-P0, CMV-P1, CMV-P2, CMV-P3, and CMV-P4. 7 host...

Cucumber Mosaic Virus Coat Protein Inside the Chloroplast of the Infected Tobacco

ZHU Shui-Fang, R.I.B. Francki

1992, 7(3): 328

The intact chloroplasts were made through protoplasts. In the SDSPAGE-silver stained gel, one extra protein band, which had the same mobility and molecular weight as the cucumber mosaic virus coat protein (CMV-cp), was found in the diseased chloroplast comparative to the health chloroplast. In the immunoblot testing, the band was highly reactive to the antiserum against the free CMV-cp, while the healthy and diseased sap treated healthy chloroplasts had no reaction. The result provides an evidence for the C...

Studies on the Viral Polypeptides and Serological Relationships of Four Strains of Rabbit Haemorrhagic Disease Virus (RHDV)

SHU Wen, LUO Jing, YANG Hua-Lou

1992, 7(3): 334

In this paper four different strains of RHDV were purified by two phases of polyethylene glyeol-dextran sodium sulfate andcentrifugation, then virions in highly pure form were obtained by using Sepharose 4B chromatography. The recovery rate of virus preparation was about 70 pereent. Serological relationships of four different strains of RHDV were studied by using immuno double diffusion, cross hemagglutination inhibition and enzyme-linked immunosorbent assay(ELISA). The results showed that serotypes are the...

Studies on Replication of Rabbit Hemorrhagic Disease Virus (RHDV) in Vitro

LUO Jing, YANG Hua-Lou, LIU Gong, YAN Yin-Fang, WANG Han-Zhong, SUN Song-Bai

1992, 7(3): 343

In this paper the characteristics of replication of RHDV in the RL and RK cells have been studied by using synchronous infective method. The cytopathology effect can be observed at 48—96 hrs after viruses infection. HA titer of viral increased 5—10 times as high as those HA titers of viruses infection at 72 hr. The sensitivity of RHDV to passage cells are different, it is most sensitive for 4—8 generation. Special fluorescence in the ceils can be seen at 18—72 hrs using immunofluorescence stain. Viruses of ...

Antigenic Relationship between Rabbit Hemorrhagic Disease Virus and Parvoviruses

LIU Dian-Zhang, XU Wei-Yan, DU Nian-Xin

1992, 7(3): 351

Antigenic relationship between rabbit hemorrhagic disease virus (RHDV) and six parvoviruses was determined with indirect enzyme-linked immunosorbent assay (indirect-ELISA), enzyme-linked immunosorbent inhibition assay (ELISIA) and hemagglutination inhibition (HI) test. The results by indirectELISA showed that the correlation coefficients between RHDV and minute virus of mice (MVM), goose parvovirus (GPV), porcine parvovirus (PPV) and mink enteritis virus (MEV) were 5.59%, 3.54%, 1.76% and 0.7%, respectively...

Restriction Enzyme Analysis and Molecular Cloning of CAV-1 DNA

BAI Li-Ping, TONG Guang-Zhi, DIAO Yong-Jun, TUN Bao-Cheng, YUE Jun-Meng, WANG Mei, YAN Shen

1992, 7(3): 357

The virulent and modified canine adenovirus type 1 (CAV-1) DNAs extracted from canine kidney cells (MDCK) 36 hrs postinfection were both analysed with restriction enzymes EcoR Ⅰ, BamH Ⅰ, Pst Ⅰ, Nsi Ⅰ, Sph Ⅰ and Hind Ⅲ. and no difference was found from the electrophoresis pattern between the two viruses. The modified CAV-1 DNA was digested completely with Pst Ⅰ and cloned into, plasmid pBluescript SK taking shotgun procedure. Through hybridization selection of photobiotin labeled CAV-1 DNA and Pst Ⅰ digestio...

Detection of Potato Spindle Tuber Viroid by Polymerase Chain Reaction(PCR)

HE Xiao-Yuan, ZHOU An-He, CHENG Zhuo-Min, WANG Li-Yang, CHEN Jian-Bei

1992, 7(3): 362

A new detection method has been developed by a new technique of polymerase chaiu reaction (PCR). According to the sequence of PSTVd, two primers were designed and synthesized with automated DNA synthesizer. The cDNA was synthesized by reverse transcription of PSTVd RNA, the cDNA was then amplified by polymerase chain reaction, the amplified products were detected by gel electrophorcsis. The results showed that the method was highly specific and the sensitivity was 0.15 pg. It was more sensitive than other m...