| Qiaoruisu is the dry grass of Coleus forskohlii(wild.),which is mainly produced in India,Sri Lanka,Nepal and other tropical or subtropical regions.It is also radically found in Yunnan,China.Known as a “magic drug”,the folks used this medicine to treat asthma,cancer and other diseases with a significant effect.Due to its high medicinal value,Hubei Furen Pharmaceutical Co.,Ltd.makes it the sovereign drug,together with Peucedanum,licorice which is developed as "Coleus Capsule",a new kind of t r d i t i o n a l C h i n e s e m e d i c i n e.T h i s p l a n t w a s a l s o transplanted from Yunnan province to Tongcheng,Hubei p r o v i n c e.L i n a l o o l i s a c o l e u s m o n o t e r p e n e,a s i s reported,it has analgesic,anxiolytic,sedative and hypnotic,anti-inflammatory,anti-tumor,anti-bacterial and other pharmacological activity,its Pharmaceutical prospects are very broad.This paper is aiming at increasing linalool content by gene technology and providing a r e l i a b l e e x p e r i m e n t a l d a t a f o r t h e s t a n d a r d i z e d cultivation of Coleus forskohlii by systematically studying on pollen morphology and pollen viability.Using Coleus forskohlii as the experimental material to extract total RNA after which a gene template was builded;design primers from gene contig by DNAMAN software and obtain full length of linalool synthase gene by RACE.Design primers from the full length gene,adding restriction enzyme cutting site Sal I and Hind III.Obtain recombinated expression vector by connecting the product to pet28 a vector,and translate the gene sequence into protein sequence.Transfer the expression vector into expression E.coli BL21(DE3).The target protein is obtained by IPTG induction,extracted and purified by Ni-Agarose Kit.Set a study on pollen morphology,pollen tube growth,and pollen viability of Coleus forskohlii,providing basis data for identification and cultivation of Coleus forskohlii. |
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