Screening of sub-clonal cell lines from IPLB-LD652Y for the mass production of Lymantria xylina multiple nucleopolyhedrovirus（LyxyMNPV）
Date Issued
2006
Date
2006
Author(s)
Lin, Kuang-Hung
DOI
zh-TW
Abstract
Seven new cell lines, IPLB-LD652Y-5, IPLB-LD652Y-7, IPLB-LD652Y-b, IPLB-LD652Y-d, IPLB-LD652Y-f, and IPLB-LD652Y-h, have been sub-cloned from the parental cell line, IPLB-LD652Y. There were four kinds of cell shape within these cell lines including round, squamous, spindle, and polymorphous shape. Among them, the round shape cells were the most preponderant. According to the virus infection results, SL-7B cannot been infected with Ly5, LyxyExp-DsRed, and LyxyExp-EGFP while the NTU-LY cell lines had high susceptibility with the same viruses infection. Moreover, the IPLB-LD652Y and subcloned cell lines showed similar results as the NTU-LY cell lines, but them showed lower susceptibility when infected with LyxyExp-DsRed, and only IPLB-LD652Y-h had lower susceptibility when infected with LyxyExp-EGFP. According to the results, we chose three sub-clonal cell lines with high production of Ly5 virus, IPLB-LD652Y-5, IPLB-LD652Y-7, and IPLB-LD652Y-f for the further comparison with the parental cell line. We can observe that the chosen cell lines have higher growth rate than the parental cell line, and the growth rate of IPLB-LD652Y-f were higher than that of IPLB-LD652Y-5.The distribution of chromosome numbers of these four cell lines were very close, between 78.02 to 87.28. In the result of isozyme, whether on EST Esterase （EST）、 Malate dehydrogenase ( MDH ) or Lactate dehydrogenase ( LDH ), the electrophoresis patterns of the chose cell lines were all the same and significant different from that of the NTU-LY1. The ITS （Internal transcribed spacer） sequences of the four cell lines, and they showed no significant difference. In the relative fluorescent unit (RFU) comparison, significant curve growth and decline with the increasing with the time post LyxyExp-EGFP inoculation were observed. Integrating the results, the IPLB-LD652Y-7 could be sifted as the highest virus susceptibility, production, and speedy growth cell line when infected with LyxyMNPV.
Subjects
舞毒蛾細胞株
木毒蛾核多角體病毒
次選殖
IPLB-LD652Y cell line
LyxyMNPV
sub-clone
Type
thesis
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