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郑鹏生, PENG-SHENG ZHENG, *†, SHOU-Rou L, *, TSUYOSHI IWASAKA, †, JIE SONG, MAN-HUA Cu, AND HAJIME SUGIMORI†
GYNECOLOGIC ONCOLOGY 58. 179-183 (1995),-0001,():
-1年11月30日
A consensus multiplex PCR rCM-PCR)technique was devel-oped to detect high-risk fHPV 16/18), low-risk(HPV 6/11), and over 40 other types of human papillomavirus(HPV), separately but simultaneously. by mixing three pairs of consensus primers in the same PCR mixture, for ene amplification. Simultaneous detection of three groups 0f HPV DNA provides valuable infor-matfon for clinical practice and this procedure is simple and convenient for routine laboratory examinations. We detected HPV DNA sequences in plasmid HPV DNA and DNA extracted from tissues of condyloma acuminata and cervical carcinoma and from exfoliated cells of the lower genital tract of healthy Chinese women living in the People’s Republic of China. We confirmed that this simple, convenient, and cost-beneficial CM-PCR technique is reliable for the detection of HPV DNA se-quences.
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郑鹏生, Peng-Sheng Zheng†, Jane Brokaw‡, and Alison A. McBride*
,-0001,():
-1年11月30日
The papillomavirus E2 protein is required for viral transcriptional regulation, DNA replication and genome segregation. We have previously shown that the E2 transactivator protein and BPV1 genomes are associated with mitotic chromosomes; E2 links the genomes to cellular chromosomes to ensure efficient segregation to daughter nuclei. The transactivation domain of the E2 protein is necessary and sufficient for association of the E2 protein with mitotic chromosomes. To determine which residues of this 200-amino-acid domain are important for chromosomal interaction,E2 proteins with amino acid substitutions in each conserved residue of the transactivation domain were tested for their ability to associate with mitotic chromosomes. Chromatin binding was assessed by using immunofluorescence on both spread and directly fixed mitotic chromosomes. E2 proteins defective in the transactivation and replication functions were unable to associate with chromosomes, and those that were competent in these functions were attached to mitotic chromosomes. However, several mutated proteins that were defective for chromosomal interaction could associate with chromosomes after treatmentwith agents that promote protein folding or when cells were incubated at lower temperatures. These results indicate that precise folding of the E2 transactivation domain is crucial for its interaction with mitotic chromosomes and that this association can be modulated.
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