Cloning of the rat progesterone receptor cDNA

Pharmaco/ogicalResearchCommun~ations, Vo~2~ Supplement~ 1988

180

CLONING OF THE RAT PROGESTERONE RECEPTOR eDNA M. G i a n n i ,

E. V e g e t o , A. Maggi

M i l a n o M o l e c u l a r Pharmacology Lab -

Inst.

Pharmacol. S c i .

-

U n i v e r s i t y o f M i l a n - P i a z z a Durante 11, ~ 0 1 3 1 M i l a n o Key words: P r o g e s t e r o n e r e c e p t o r ,

In order

to

receptor RNA

isolate

(PR),

purified

oligo(dt)

from r a t

library

(2),

utilizing

as

conditions

uterus.

The eDNA was s y n t h e s i z e d v i a

were

the screening of probe the

chicken

to

rat

uterus

with

poly(A)+

the p u t a t i v e

was

the

performed The

heterologous capability

RNA

and

spotted

to onto

The p r i m a r y s c r e e n i n g was performed

clones of

the

amplified

positives

isolated

screening.

Among t h e p o s i t i v e s

kb i n s e r t

Since

human ~'PPR eDNA.

hybridization

filters.

I0 ~

chicken

the l i b r a r y or

(l>.

the chicken

e s t a b l i s h e d by e x a m i n i n g t h e i r

nitrocellulose about

homology has been d e s c r i b e d i n

a

for

hybridize

the

progesterone

was p r e p a r e d from p o l y ( A ) +

and hormone b i n d i n g domains o f

human PR,

a 2.1

the r a t

p r i m i n g and f o l l o w i n g G u b l e r ' s p r o t o c o l

DNA-binding

on

the eDNA c o d i n g f o r

a ~gt10

a high degree o f

probes

eDNA c l o n i n g

underwent

secondary isolated,

which h y b r i d i z e s w i t h

eDNA, but

not

library.

with

e s t r o g e o r e c e p t o r and g l u c o c o r t i c o i d

20

tertiary

one c l o n e c o n t a i n s

the 5'

the 5'

and

The

end (500 bp) o f

end

of

the

rat

receptor.

REFERENCES: 1. 2.

Gubler U. and Hoffman B . J .

(1983) Gene 2 5 : 2 6 3 - 2 6 9

Misrahi M., et al. (1987) B i o c h e m i c a l and B i o p h y s i c a l Research Communications 1 4 3 : 7 4 0 - 7 4 8 .