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Mutation and selection: Shaping the...

O'Neil, Patrick Kevin.

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Mutation and selection: Shaping the HIV-1 genome.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Mutation and selection: Shaping the HIV-1 genome./
作者:	O'Neil, Patrick Kevin.
面頁冊數:	168 p.
附註:	Source: Dissertation Abstracts International, Volume: 64-02, Section: B, page: 0698.
Contained By:	Dissertation Abstracts International64-02B.
標題:	Chemistry, Biochemistry. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3082863

Mutation and selection: Shaping the HIV-1 genome.
O'Neil, Patrick Kevin.

Mutation and selection: Shaping the HIV-1 genome. - 168 p.

Source: Dissertation Abstracts International, Volume: 64-02, Section: B, page: 0698.

Thesis (Ph.D.)--The University of Utah, 2003.

This thesis details research on the rate, nature and consequences of mutation in human immunodeficiency virus type 1 (HIV-1). The rate and nature of HIV mutation was determined by examining proviral long terminal repeats (LTRs) for mutations after a single cycle of viral replication in cell culture. The LTRs were selected because, as a consequence of the way that retroviruses replicate, they can provide information as to when during viral replication mutations occurred. The LTRs of 215 unique proviruses were examined by PCR fragment size screening, single strand conformation polymorphism (SSCP), and simple sequencing. Besides determining a mutation rate for HIV this research reports the first known example of HIV transducing host cell DNA. Ultimately, a total of 22 mutations of various types were detected including base substitutions, deletions, insertions, and rearrangements. Base substitutions predominated with the majority of these being N to A changes. The overall mutation rate was quite high at 8.5 × 10<super>−5</super> per basepair per replication cycle, a few-fold higher than previous reports from single cycle assays using foreign reporter genes.Subjects--Topical Terms:

1017722
Chemistry, Biochemistry.

Mutation and selection: Shaping the HIV-1 genome.
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502 $a Thesis (Ph.D.)--The University of Utah, 2003.
520 $a This thesis details research on the rate, nature and consequences of mutation in human immunodeficiency virus type 1 (HIV-1). The rate and nature of HIV mutation was determined by examining proviral long terminal repeats (LTRs) for mutations after a single cycle of viral replication in cell culture. The LTRs were selected because, as a consequence of the way that retroviruses replicate, they can provide information as to when during viral replication mutations occurred. The LTRs of 215 unique proviruses were examined by PCR fragment size screening, single strand conformation polymorphism (SSCP), and simple sequencing. Besides determining a mutation rate for HIV this research reports the first known example of HIV transducing host cell DNA. Ultimately, a total of 22 mutations of various types were detected including base substitutions, deletions, insertions, and rearrangements. Base substitutions predominated with the majority of these being N to A changes. The overall mutation rate was quite high at 8.5 × 10<super>−5</super> per basepair per replication cycle, a few-fold higher than previous reports from single cycle assays using foreign reporter genes.
520 $a This thesis also details preliminary research to determine the import of a striking characteristic of HIV and other lentiviruses: a marked A-bias in the nucleotide makeup of the viral genome. In an effort to determine what biological significance this bias holds for the virus, competition experiments were conducted in which a series of HIV-1 variants containing “silent” nucleotide changes reducing the A-bias within a localized region of the viral genome were competed against wildtype virus over repeated passages in cell culture. After each passage, a sample of cell culture supernatant containing infectious virus was taken to determine the makeup of the viral population. Changes in the proportion of wildtype to variant over time can be used to determine the relative fitness of variants vs. wildtype virus. The preliminary data from two different cell lines indicated no obvious fitness effects due to the nucleotide changes. Difficulties were encountered within the experimental system used, however, that can be addressed in the future studies.
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