Construction of cDNA libraries from Chinese shrimp Fenneropenaeus chinensis

Six cDNA libraries of blood,eyestalks,ovary and cephalathorax of female,male,triploid Chinese shrimp(Fenneropenaeus chinensis) were constructed using the cDNA synthesis and ZAP express kit (Stratagene).Total RNA was extracted with guanidine thiocyanate/phenol/chloroform.Synthesized cDNA was ligated with the E coR I adapter and phosphory lated.After EcoR I adapters were digested and eliminated with Sephrose-2Bspun column,the cDNA fragments that had more than 400 bp were collected and ligated with the ZAP express vector.And then the ligated cDNAs were packed and incubated with XL blue MRF.The content of each library is 0.78×10⁶ (blood),0.60×10⁶(eyestalks),0.36×10⁶(ovary),1.3×10⁶(female),0.20×10⁶(male) and 1.2×10⁶(triploid),respectively.In order to detect the quality of these libraries,the inserted fragments were am plified with the T3 and T7 primers.The size of the inserted fragments was 500~2500 bp.Moreover,antimicrobial peptide and-actingenes were cloned from the libraries.All these results indicate that high quality cDN A libraries were obtained.These cDNA libraries provide avery powerful tool to find new functional genes,analyze ESTs,and prepare genechips and so on.