[Screening and identification of genes trans-regulated by a novel HbeAg interacting protein AK026018 with microarray assay].
BACKGROUND
To investigate the biological functions of a novel hepatitis B virus e antigen (HbeAg) interacting protein AK026018, and to use cDNA microarray technique to screen genes regulated by the protein.


METHODS
The AK026018 coding DNA fragment was amplified by reverse transcription polymerase chain reaction (RT-PCR) technique from HepG2 cell. The expressive vector of pcDNA3.1-AK was constructed by routine molecular biological methods. The HepG2 cells were transfected with pcDNA3.1 and pcDNA3.1-AK, respectively by using lipofectamine. The total RNA was isolated and reverse transcribed. The cDNA of each sample was subjected to microarray screening with 8,464 cDNA probes and analyzed by bioinformatics.


RESULTS
The expressive vector was constructed and confirmed by DNA sequencing analysis and restriction enzyme digestion. High quality mRNA and cDNA of transfected HepG2 cells had been prepared and successful microarray screening conducted. From the scanning results, there were 122 differential expression genes, of which 36 genes were down-regulated, and 16 genes were up-regulated.


CONCLUSIONS
Microarray technique was successfully used to screen the genes trans-regulated by AK026018. The expression of AK026018 protein affects the expression spectrum of HepG2 cells.
Li
Bo-an
BA
Department of Immunology, Institute of Infectious Disease, No. 302 Hospital, Beijing 100039, China.
Hou
Jun
J
He
Wei-ping
WP
Qi
Yang
Y
Chi
Shu-ping
SP
Zhao
Jun
J
Cheng
Yun
Y
D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D045744:Cell Line, Tumor; D019295:Computational Biology; D015972:Gene Expression Regulation, Neoplastic; D006513:Hepatitis B e Antigens / Q000378:metabolism*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D002352:Carrier Proteins; D006513:Hepatitis B e Antigens