Disease relevance of pol

• Two methods for the extraction of total NSC DNA from up to 2 ml of non-extended semen were employed; however, no BIV pol fragment was amplified from these DNA preparations [1].
• We have produced sequence data representing 598 bp of the pol gene, amplified by PCR from viral cDNA using broadly reactive universal primers for retroviruses and more specific genus-reactive primers for lentiviruses [2].
• Blood samples were collected following delivery of calves, the peripheral blood leukocytes were purified from these samples, and the leukocyte DNA was used in polymerase chain reactions targeting the pol gene region of the BIV provirus [3].
• Our present studies show that this virus encodes a reverse transcriptase (RT) with Mg2+ cation preference, replicates and induces syncytia in a variety of embryonic bovine tissues in vitro, and has a morphology most similar to the human immunodeficiency virus (HIV) [4].
• As with the feline representative, BIV also was found to possess a lentivirus morphology and to encode a reverse transcriptase with Mg++ preference; it replicates and induces syncytia in a variety of embryonic bovine tissues in vitro [5].

High impact information on pol

• To resolve the phylogenetic relationship of this virus, proviral molecular clones were derived and used to determine the nucleotide sequence of the highly conserved RT domain [4].
• In order to minimize the sequence homologies, we recoded gag/pol with codon usage optimized for expression in human cells in this report [6].
• The ribonuclease-H cleavage pattern suggests that the spatial distance between the polymerase and ribonuclease-H active sites of the two BIV RT isoforms equals 20 nt, unlike the 17 nt distance observed in almost all other RTs [7].
• We examined the genetic diversity and natural selection of a segment of the BIV pol gene amplified from the leukocyte DNA of naturally infected cattle [8].
• Reverse transcriptase activity was detected in virus purified by sucrose gradient centrifugation [9].

Biological context of pol

• Primers were designed to amplify 2 separate regions of the pol and env segments of the BIV genome [10].
• Sequence analyses of the polymerase chain reaction pol gene product showed 92.6 and 93.6% homology to the published nucleotide sequence of BIV R29-127, a molecular clone derived from BIV R29 [11].
• Two primer pairs located in the gag and pol regions of the viral genome allowed to detect the viral genomic DNA by PCR, as well as the unspliced genomic viral RNA transcript, by RT-PCR [12].
• Amino acid sequences of these Zambian pol gene products showed 98.0 to 100% homology to the American strain BIV R29, 97.0 to 99.0% to Japanese BIV isolates, and divergence ranged from 0.0 to 2.0% among Zambian BIV isolates [13].

Anatomical context of pol

• PROCEDURE: Polymerase chain reaction testing that used 3 sets of primer pairs targeting pol and env regions of the BIV proviral genome was performed on DNA extracted from semen leukocytes, spermatozoa, and blood leukocytes from each bull [14].
• PCR, Southern blot hybridisation, cell culture and reverse transcriptase assays were used to demonstrate the presence of BIV proviral DNA and the production of infectious virus in CD2+, WC1+, B cells and monocytes during the acute stages of infection [15].

Analytical, diagnostic and therapeutic context of pol

• PCR analysis of MNC DNA, using BIV pol gene primers, detected virus in all three of the experimentally infected bulls from DPI 9 until the termination of the experiment at DPI 98 [1].
• Two pairs of oligonucleotide primers directed to sequences within the coding regions of the gag and pol genes generated the expected PCR products from molecular BIV clones and from DNA of BIV-infected cell cultures but not from DNA of uninfected cultures [16].
• Microinjection of purified DNA from BIV clone 106 or 127 into susceptible bovine cells produces virus-specific cytopathic effects, including syncytium induction, supernatant reverse transcriptase activity, and infectious virus particle formation, similar to the effects produced by parental virus stock [17].
• Bovine immunodeficiency virus (BIV) was purified by isodensity centrifugation; viral activities were monitored in gradient fractions using the reverse transcriptase assay and a p26-specific monoclonal antibody ELISA [18].

References