Human adenovirus type 41 possesses different amount of short and long fibers in the virion.
To determine the ratio of short fiber (sfiber) to long fiber (lfiber) in human adenovirus type 41 (HAdV-41) virions, sfiber and lfiber were expressed in E. coli, quantified, and used as loading standards in Western blot. Densitometric analyses of the standard and target bands indicated that the ratio of sfiber to lfiber in virions was 5.7±0.7. Sfiber-deleted HAdV-41, HAdV-41-DSF-GFP, was constructed, and Western blot analysis showed that the amount of lfiber in HAdV-41-DSF-GFP was about 7.3±1.9 times of that in HAdV-41 virions, confirming a ratio of approximate 6 for sfiber to lfiber in HAdV-41. In HAdV-41-infected cells, mRNAs of the sfiber and lfiber were comparable in quantity, while the expression at protein level was significantly different. Our results suggested an unequal number of short and long fibers, which might result from their differential protein expression during HAdV-41 packaging. The method used here could be extended to quantify other trace proteins.
National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 100 Ying Xin Jie, Xuan Wu Qu, Beijing 100052, PR China.
Jing-Dong
JD
Song
Xue-Li
XL
Liu
Duo-Ling
DL
Chen
Xiao-Hui
XH
Zou
Min
M
Wang
Jian-Guo
JG
Qu
Zhuo-Zhuang
ZZ
Lu
Tao
T
Hung
D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002213:Capsid / Q000737:chemistry / Q000378:metabolism; D036022:Capsid Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014771:Virion / Q000737:chemistry* / Q000378:metabolism
D036022:Capsid Proteins; C018315:hexon capsid protein, Adenovirus