Screening of differentially expressed genes in placentas with hepatitis B virus infection by suppression subtractive hybridization technique

• Published in: Chung-hua fu chʿan kʿo tsa chih Vol. 42; no. 2; p. 76
• Main Authors: Bai, Gui-Qin, Yue, Ya-Fei, Zhang, Shu-Lin, Cheng, Jun, Liu, Yan, Li, Shu-Hong, Zhang, Xin-E
• Format: Journal Article
• Language: Chinese
• Published: China 01.02.2007
• Subjects: Cloning, Molecular; DNA, Complementary - genetics; Female; Gene Expression; Gene Library; Genes, Viral - genetics; Hepatitis B virus - enzymology; Hepatitis B virus - genetics; Humans; Molecular Sequence Data; Nucleic Acid Hybridization; Phosphatidylinositol 3-Kinases - genetics; Phosphatidylinositol 3-Kinases - metabolism; Placenta - enzymology; Placenta - virology; Pregnancy; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism
• ISSN: 0529-567X

To screen differentially expressed genes in placentas with hepatitis B virus (HBV) infection and to discuss the molecular mechanism of HBV intrauterine infection. Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group. The suppression subtractive hybridization (SSH) technique was used. Total RNAs of placenta tissue of the study group were mixed as the tester, and total RNAs of placenta tissue of the control group were mixed as the driver. A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems. Amplifications of the library were carried out with E. coli strain DH5alpha by reverse spot hybridization. RT-PCR confirmed that phosphatidylinositol 3-kinase (PI3K) was up-regulated in placenta tissue with HBV infection. Colony PCR showed that the clones contained 200 - 1000 bp inserts. Thirty five clones were c