APPLICATION OF RECOMBINANT DNA TECHNOLOGY IN

PHARMACEUTICAL PRODUCTIONS

BACKGROUND:

Catharanthus roseus, or commonly called Periwinkle, is a valuable

medicinal plant known for its production of a variety of Terpenoid Indole
Alkaloids (TIAs), such as vindoline, vinblastine, etc. These TIAs are essential
components in the production of drugs. Hence, in order to improve the
production of alkaloids, Catharanthus roseus is genetically transformed,
definitely, with the help of recombinant DNA technology.

HOW IT IS DONE

To develop a genetic transformation method for C. roseus, bacterial genes

were added to the plant. The bacterial DNA may come from either
Agrobacterium rhizogenes or Agrobacterium tumefaciens. However, the
phenotypes of periwinkle plants transformed by A. rhizogenes are abnormal,
having shortened internodes, wrinkled leaves and abundant root mass, thus, it
is not suitable for the production of alkaloids. Hence, A. tumefaciens can be
used to develop transgenic C. roseus plant. During the process, the
association parameters were systemically investigated and optimized,
including the concentration of Agrobacterium and acetosyringone, genetic
transformation and co-cultivation duration, plant regeneration and analysis of
putative transformants.

PROCESS

Seeds of C. roseus are sterilized and germinated.For

transformation, Agrobacterium tumefaciens strain EHA105 harbors a
binary vector－ pCAMBIA2301, which contains β-glucuronidase (GUS)
gene and a selectable marker neomycin phosphotransferase II gene
(NTPII). The explants of C. roseus is introduced for co-cultivation
for about 1-3 days in a dark place with a temperature of 28℃. After
co-cultivation, the explants were transferred into an induction
medium for plant regeneration. After the integration of GUS gene into
the C. roseus plant, the expression of GUS gene was examined after 5
days of co-cultivation. In order to validate the established
transformation events/system, analysis of real-time PCR and HPLC is
done, and southern blot analysis is carried out.

IMMEDIATE BENEFITS?
 The enhanced production of Terpenoid Indole Alkaloids (TIAs) in modified
periwinkle plants are usually added to drugs for cancer treatments. For
instance, the alkaloid vinblastine is an essential component of drugs to treat
patients with Hodgkin’s lymphoma. In addition, the alkaloid vindoline is also an
essential component of drugs to reduce hyperlipidemia and renal failures.
Vindoline can also reduce the oxidative stress in diabetic hearts. Hence, the
modified periwinkle plant can help to treat various diseases, such as cancer,
cardiovascular diseases and kidney diseases.

REFERENCES:

Wang, Q., Xing, S., Pan, Q. et al. (2012). Development of Efficient

Catharanthus Roseus Regeneration and Transformation System Using
Agrobacterium Tumefaciens and Hypocotyls as Explants. BMC
Biotechnol 12, 34.
https://bmcbiotechnol.biomedcentral.com/articles/10.1186/1472-6750-12-
34#citeas

Oguntibeju O., Aboua Y., & Goboza M. (2019). Vindoline—A Natural Product
from Catharanthus Roseus Reduces Hyperlipidemia and Renal
Pathophysiology in Experimental Type 2 Diabetes. Biomedicines, 7(3), 59.
https://www.mdpi.com/2227-9059/7/3/59/htm

Serafica J.P., Pawilen G., Caslib B., & Alata E.J. (2018). Genetically Modified
Organisms. Science, Technology, and Society. (1st ed., pp. 134-147).
REX Book Store.