I was trying to express the protein of sri lankan cassava mosaic virus coat protein gene which was cloned into pET28A+ expression vector and transformed into BL21(DE3) cells. Insead of getting the desired band size I am getting large band size after IPTG induction (instead of getting 35 kDa, i am getting around 120 kDa protein band size induced in SDS PAGE ). Can anyone help me to solve this issue?
Hello,
Did you get the correct band size after PCR amplification?
Which restriction sites did you opt for?
Did you go for sequencing of the clone you used before the expression of protein?
It will help in understanding the situation better. Also, can you show me the gel picture?
Can you share with us the sds-page immage?
How did you prepare the samples vedi te rum the sds page ? ( eg Reducing agent - booiling)
Ciao
Manuele
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