Method of providing a hybrid polypeptide exhibiting an...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C435S029000, C435S091200, C435S069100, C435S455000, C435S471000, C435S476000

Reexamination Certificate

active

06270968

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to a method of providing novel DNA sequences encoding a polypeptide with an activity of interest, novel DNA sequences provided according to the method of the invention, polypeptides with an activity of interest encoded by novel DNA sequences of the invention.
BACKGROUND OF THE INVENTION
The advent of recombinant DNA techniques has made it possible to select single protein components with interesting properties and produce them on a large scale. This represents an improvement over the previously employed production process using microorganisms isolated from nature and producing a mixture of proteins which would either be used as such or separated after the production step.
Since the traditional methods were rather time-consuming, more rapid and less cumbersome methods were developed.
A such technique is described in WO 93/11249 (Novo Nordisk A/S).
The method described in WO 93/11249 comprises the steps of:
a) cloning, in suitable vectors, a DNA library from an organism suspected of producing one or more proteins of interest;
b) transforming suitable yeast host cells with said vectors;
c) culturing the host cells under suitable conditions to express any protein of interest encoding by a clone in the DNA library; and
d) screening for positive clones by determining any activity of a protein expressed in step c).
According to this method it is necessary to prepare a DNA library, comprising complete genes encoding polypeptides with activities of interest. Such a library has traditionally been made on mRNA isolated from micro-organisms which has been cultivated and isolated.
As it is only possible with known methods to cultivate about 2% of the microorganisms known today (i.e. cultivable microorganisms), genes encoding polypeptides from a huge number of microorganisms (i.e. un-cultivable microorganisms) are generally difficult to identify and clone on the basis of screening technologies used today, such as the above mentioned.
SUMMARY OF THE INVENTION
It is the object of the present invention to provide a method for providing a novel DNA sequence encoding a polypeptide with an activity of interest from micro-organisms without having to cultivate and isolate said micro-organisms.
In the first aspect the invention relates to a method of providing novel DNA sequences encoding a polypeptide with an activity of interest, comprising the following steps:
i) PCR amplification of said DNA with PCR primers with homology to (a) known gene(s) encoding a polypeptide with an activity of interest,
ii) linking the obtained PCR product to a 5′ structural gene sequence and a 3′ structural gene sequence,
iii) expressing said resulting hybrid DNA sequence,
iv) screening for hybrid DNA sequences encoding a polypeptide with said activity of interest or related activity,
v) isolating the hybrid DNA sequence identified in step iv)
Further, the invention also relates novel DNA sequences provided according to the method of the invention and polypeptides with an activity of interest encoded by said novel DNA sequences of the invention.


REFERENCES:
patent: 5457046 (1995-10-01), Woldike et al.
patent: 5512478 (1996-04-01), Orser et al.
patent: 5780225 (1998-07-01), Wigler et al.
patent: 5849491 (1998-12-01), Radomski et al.
patent: 0 368 684 (1990-05-01), None
patent: WO 91/17244 (1991-11-01), None
patent: WO 91/17243 (1991-11-01), None
patent: WO 93/11249 (1993-06-01), None
patent: WO 94/07998 (1994-04-01), None
patent: WO 96/29397 (1996-09-01), None
Okuta et al., Gene 212, 221-228 (1998).*
Heussler et al., Mol. Biochem. Parasit. 64, 11-23 (1994).*
Repaske et al., J. Biol. Chem. 267(26), 18683-18688 (1992).*
Sarantopoulos et al., Journal of Immunology, vol. 152, No. 11, pp. 5344-5351 (Jun. 1, 1994).
Henrissat et al., Journal Biochem., vol. 280, pp. 309-316 (1991).
Smalla et al., Journal of Applied Bacteriol., vol. 74, pp. 78-85 (1993).
Stein et al., Journal Bacteriol., vol. 178, No. 3, pp. 591-599 (Feb. 1996).
Embleton et al., Nucleic Acids Research, vol. 20, No. 15, pp. 3831-3837 (1992).

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