Claims
- 1. A process for the preparation of DNA encoding the seed specific AmA1 protein from Amaranthus which comprises:
- i) isolating messenger RNA from Amaranthus by conventional methods;
- ii) preparing from mRNA double stranded cDNA with reverse transcriptase using oligo (dT) primers;
- iii) combining the double stranded cDNA with an expression vector;
- iv) isolating from the resulting recombinant vectors those encoding said seed specific protein by conventional methods; and
- v) sequencing the DNA encoding said seed specific protein.
- 2. A process as claimed in claim 1 wherein the isolation of messenger RNA from total RNA of Amaranthus seeds is done by oligo (dT) cellulose chromatography.
- 3. A process as claimed in claim 1 wherein the synthesis of the double stranded cDNA is effected by reverse transcriptase using poly(A).sup.+ RNA as template and oligo (dT) 12-18 as primer to obtain full length cDNA.
- 4. A process as claimed in claim 1 wherein the expression vector used is a disarmed bacterial virus.
- 5. A process as claimed in claim 1 wherein the double stranded cDNA is grown as phage plaques and recombinant proteins expressed by phages are transferred to filters and the recombinant clone identified using antibodies raised against said seed specific Ama1 protein.
Parent Case Info
This is a division of application Ser. No. 08/158,270 filed Nov. 29, 1993 now U.S. Pat. No. 5,670,635.
Non-Patent Literature Citations (2)
Entry |
Promega Protocols and applizations Guide, Second Edition, Mar. 1991. Promega Corporation, Madison WI. |
Marcone, M.F, and R.Y. Yada. 1991. Agriz. Biol. Chem. vol. 55, No. 9, pp. 2281-2289. |
Divisions (1)
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Number |
Date |
Country |
Parent |
158270 |
Nov 1993 |
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