Cloning And Expression System Of CrtE,CrtI,CrtYB Gene From Phaffia Rhodozyma

P-carotene,one of the carotenoids,is an orange fat soluble substance.?-carotene is widely used in food,feed,cosmetics and medical fields due to its good geographic properties.The market demand is becoming more and more vigorous.The main source of ?-carotene is chemical synthesis and natural extraction.Natural extracted ?-carotene is more safe and widely used.This paper extracts the target genes of crt E,crtI,crt YB genes from Phaffia rhodozyma,the target gene was cloned into GAP-crtE-CYC1,GAP-crtI-CYC1,GAP-crtYB-CYC1 box construction and expression of p426GPD in.The expression box was further integrated into the pRS406 expression vector to construct the expression vector.The results of this experiment is as follows:by using Kpnl and Clal double enzyme digestion,the GAP-crtE-CYC1 vector was added into pRS406 expression vector,and the expression vector pRS406-crtE was obtained.The pRS406-crtE-crtYB expression vector was obtained after pRS406-crtE was cut off at the end of the EcoRI enzyme and was connected with the target fragment GAP-crtYB-CYC1.Finally,the final expression vector of pRS406-crtE-crtYB-crtIv btained by using NotI enzyme to cut the ends of GAP-crtI-CYC1 fragments.