[Construction and screening of suppression subtractive hybridization library of renal cell carcinoma].
OBJECTIVE
To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC).


METHODS
Poly A(+) RNA was isolated from RCC lines 786-O (tester) and renal cell (RC) lines HK-2 (driver), respectively. SSH procedure was performed according to the protocol of the PCR-Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced.


RESULTS
The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes.


CONCLUSIONS
The quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC.
Zhang
Yong
Y
Institute of Urology, First Hospital, Peking University, Beijing 100034, China.
Ai
Jun-kui
JK
Zhang
Zhi-wen
ZW
Ye
Xiong-jun
XJ
Zhu
Hong-jian
HJ
Xin
Dian-qi
DQ
Liang
Li-li
LL
Na
Yan-qun
YQ
Guo
Ying-lu
YL
D018262:Adenocarcinoma, Clear Cell / Q000235:genetics*; D045744:Cell Line, Tumor; D015723:Gene Library; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods*