Figure 1.

Cloning and expression of light meromyosin in E. histolytica. (A) The 3′ end of the gene encoding the myosin heavy chain was amplified by PCR. Oligonucleotides were designed to incorporate an epitope tag at the amino terminus of the expressed LMM. The generated DNA fragment was cloned into the ExEhNeo vector and introduced into the amoebae by electroporation. (B) Immunoblot of proteins obtained from the transfectant cells revealed by the anti-myosin II and by the anti-VSV-G antibodies. Myosin is at 250 kDa, and VSV-LMM is at 71 kDa. Lane 1, LMM20; lane 2, ExEh20, lane 3, LMM30; lane 4, ExEh30. Analysis shows that the LMM protein is expressed and correctly tagged.