摘要:

Antisense oligodeoxynucleotide (ODN), which are directed against the splice acceptor site of exon II of the regulatory gene tat of the human immunodeficiency virus type 1 (HIV-1), have been described. These 20-mer ODN’s displayed moderate anti-HIV activity in vitro. Using the same antisense ODN (termed ODN-2), which was additionally modified and protected both at the 3’- and the 5’-terminus by two phosphorothioate internucleotide linkages, a strong anti-HIV activity (EC50: 2.7 µg/ml) could be measured in the HIV-1/CEM- and HIV-l/HeLa-T4+ cell system. The analogous ODNs which were protected only at one end were either inactive (up to 10 µg/ml) or displayed a low antiviral activity. Time kinetic studies revealed that the antisense ODN-2 reduced the release of HIV-1 already after an incubation time of 1 h. By applying SI nuclease protection procedures, it could be established that the antisense ODN-2 inhibited splicing of high molecular weight transcript to the 2-kb tat mRNA in HIV-1-infected CEM cells. Transfection experiments with pU3R-III chloramphenicol acetyltransferase expression vector in HeLa-T4+ cells revealed that the antisense ODN-2 blocked the Tat protein-mediated transactivation process. In co-transfection experiments using pSV2tat72 or scrape loading studies with purified Tat, the transactivation was restored. These data indicate that the selected antisense ODN-2 displays its anti-HIV effect by blocking the splicing process leading to the functional 2-kb tat <

ISSN:0300-5526    

DOI:10.1159/000150233

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

In contrast to the strong association of human papillomavirus (HPV) type 16 with genital malignancies, HPV 6 has been found essentially in benign genital lesions. In these studies we show that HPV type 6 and 16 DNAs behave differently also in their ability to transform NIH 3T3 cells in cooperation with the carcinogen N-methyl-N’-nitro-N-nitrosoguanidine (MNNG). Although we could show that both HPV-6- and HPV-16-transfected genomes were integrated and expressed in NIH 3T3 cells, only the NIH 3T3 cells which contained the HPV 16 genome became fully transformed after MNNG treatment, as assessed by their ability to form colonies in soft agar and to induce tumors in nude mice. NIH 3T3 cells containing the HPV 6 genome and treated with MNNG did not show this potential. Furthermore, we could detect an increased expression of the E7 gene of HPV 16 in the carcinogen-treated cells containing the HPV 16 genome. These studies indicate that the presence of the HPV 16 genome specifically is also an essential step to in vitro cellular transformatio

ISSN:0300-5526    

DOI:10.1159/000150234

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Ad7h is a newly identified genome type associated with severe lower acute respiratory infections and has so far been isolated only in South America. To obtain a clue to its possible origin, the degree of restriction enzyme site homology between adenovirus genome type 7h and those representative of the three described genomic clusters (GC) for serotype 7 was studied by analysis of pairwise comigrating DNA restriction fragments (PCRF) after digestion with BamHI, BglI, BglII, BstEll, EcoRl, Hindlll, Hpal, SalI, Smal, Xbal, and Xho 90) were observed with any of the genome types grouped within GC3 or with Ad7g (the only member of GC2), so since Ad7h seems to be related to both members of GC2 and GC3, it could equally be considered to represent a new cluster or to end up grouped in either GC2 or GC3, depending on the results of further analysis.

ISSN:0300-5526    

DOI:10.1159/000150235

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

The genome of a papillomavirus isolated from a cutaneous lesion on the head of an African grey parrot (Psittacus erithacus timneh) was cloned into pBR322, and a restriction map was prepared. Several short portions of the DNA were sequenced allowing the genome to be aligned with HPV-1a. In Southern blot hybridizations, under conditions of low and medium stringency (Tm–40and –33 °), but not at a higher stringency, this viral DNA annealed weakly with only 1 of 17 mammalian papillomavirus genomes tested. Furthermore, PePV DNA hybridized with the DNA from the European chaffinch only at low stringency, indicating that it represents a unique avian papillomav

ISSN:0300-5526    

DOI:10.1159/000150236

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Ethidium bromide staining of electrophoretically separated ssRNAs and dsRNAs as well as northern blot analyses with cDNA clones suggested that the genome of the Ahlum serotype of beet soil-borne virus (BSBV) consists of two major ssRNA species of approximately 3.6 and 3.2 kb, respectively, and possibly a minor ssRNA of approximately 6.0 kb. A few of our clones hybridized with both the 3.6-kb and the 3.2-kb RNAs, the majority of the clones, however, hybridized only with the 3.2-kb RNA. The 3.2-kb RNA is, therefore, apparently not a degradation product or a partially deleted form of the 3.6-kb RNA. None of our clones hybridized with the faint band(s) of the 6.0-kb ssRNA(s) which was produced by RNA extracts of some of our virus preparations. A fourth ssRNA of approximately 3.0 kb, which hybridized with the same clones as the 3.2-kb RNA, was found at relatively high concentrations in RNA extracts from purified virus, but not in total RNA extracts from leaves. Its origin is unknown. It is apparently not derived from the 3.2-kb RNA by the loss of a VPg or a poly(A) tail. Hybridization tests with 32P-labelled poly(dT) suggested that none of the RNAs of BSBV is polyadenylated. With respect to size and the lack of a poly(A) tail the RNAs of BSBV are more similar to those of definitive furoviruses than to those of beet necrotic yellow vein virus which is only a possible member of the furovirus group and has RNAs which readily hybridized with poly(dT).

ISSN:0300-5526    

DOI:10.1159/000150237

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Two different DNA fragments, deriving from the P22-related bacteriophage LP7 and from the right IS element of transposon Tn10, have been identified which were recognized by the P22-packaging apparatus as pac-like signals. Their nucleotide sequences were compared with the known sequence of P22 gene 3 which contains the P22 pac site. Both sequences show similarity to a particular segment of P22 DNA close to a region identified earlier as part of pαc.

ISSN:0300-5526    

DOI:10.1159/000150238

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

The adeno-associated virus 2 (AAV) genome can be rescued either from a recombinant plasmid upon transfection into human cells or from cells latently infected with AAV, following subsequent infection with adenovirus. Using human diploid fibroblasts as a model for a natural AAV infection, we observed increased efficiency of rescue of the AAV genome in these cells as they traversed their limited proliferative life span in vitro. The efficiency of rescue correlated well with the augmented nuclease activity in these cells. Furthermore, rescue of the AAV genome, either from a recombinant plasmid or from the chromosomal DNA, was more efficient in cells from a patient with Bloom’s syndrome, a rare autosomal recessive disease associated with increased chromosomal breakage due to DNA ligase I deficiency, as compared with normal human diploid fibroblasts. These studies suggest that alterations in DNA-modifying enzymes may play a role in rescue of the AAV genome in human cell

ISSN:0300-5526    

DOI:10.1159/000150239

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

The majority, if not the entire life cycle, of the large dsDNA-containing algal virus PBCV-1 occurs in localized regions in the cytoplasm. Thirteen drugs that disrupt the cytoskeleton had no effect on PBCV-1 replication at concentrations which inhibited host growth. Therefore, host cytoskeletal elements do not appear to be important in PBCV-1 morphogenesis.

ISSN:0300-5526    

DOI:10.1159/000150240

出版商:S. Karger AG    

年代:1992

数据来源: Karger