[Construction of forward and reverse subtracted cDNA libraries between muscle tissue of Meishan and Landrace pigs].
Using suppression subtractive hybridization (SSH) technique, forward and reverse subtracted cDNA libraries were constructed between Longissimus muscles from Meishan and Landrace pigs. A housekeeping gene, G3PDH, was used to estimate the efficiency of subtractive cDNA. In two cDNA libraries, G3PDH was subtracted very efficiently at appropriate 2(10) and 2(5) folds, respectively, indicating that some differentially expressed genes were also enriched at the same folds and the two subtractive cDNA libraries were very successful. A total of 709 and 673 positive clones were isolated from forward and reverse subtracted cDNA libraries, respectively. Analysis of PCR showed that most of all plasmids in the clones contained 150-750 bp inserts. The construction of subtractive cDNA libraries between muscle tissue from different pig breeds laid solid foundations for isolating and identifying the genes determining muscle growth and meat quality, which will be important to understand the mechanism of muscle growth, determination of meat quality and practice of molecular breeding.
College of Animal Science and Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China.
De-Quan
DQ
Xu
Yi-Bing
YB
Zhang
Yuan-Zhu
YZ
Xiong
Jian-Fang
JF
Gui
Si-Wen
SW
Jiang
Yu-Hong
YH
Su
D000818:Animals; D005260:Female; D015723:Gene Library; D009132:Muscles / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification; D013552:Swine / Q000235:genetics*
D012313:RNA