Diagnostic test for vitamin B12

• US 7,932,369 B2
• Filed: 07/08/2009
• Issued: 04/26/2011
• Est. Priority Date: 02/07/2005
• Status: Expired due to Fees

• Current Assignee: John F. Uher, Ramani S. Wonderling
• Original Assignee: John F. Uher, Ramani S. Wonderling
• Inventors: Wonderling, Ramani S., Uher, John F.
• Primary Examiner(s): Noakes; Suzanne M
• Assistant Examiner(s): Lee; Jae W
• Application Number:
  US12/499,625
• Publication Number:
  US 20090305348A1
• Time in Patent Office: 657 Days
• Field of Search: None
• US Class Current: 536/23.1
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: G01N 33/82 involving vitamins or their receptors
• CPC Class Codes: Y10T 436/11 Automated chemical analysis

Abstract

An isolated nucleotide sequence or fragment thereof encoding the porcine intrinsic factor, wherein the porcine intrinsic factor comprises an amino acid sequence having at least 85% amino acid sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 6, and SEQ ID NO: 9. The invention also encompasses an isolated nucleic acid sequence or fragment thereof comprising, or complementary to, a nucleotide sequence having at least 85% nucleotide sequence identity to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 4, and SEQ ID NO: 7. The porcine intrinsic factor can be use is an assay to determine the quantity of vitamin B₁₂ in a biological sample.

Claims

1. An isolated nucleic acid comprising a nucleotide sequence encoding porcine intrinsic factor, wherein the amino acid sequence of said porcine intrinsic factor has at least 85% identity to SEQ ID NO:

9.

2. The nucleotide sequence of claim 1 wherein said nucleotide sequence is isolated from Sus genus.

3. The nucleotide sequence of claim 2 wherein said Sus genus is Sus scrofa.

4. An isolated nucleic acid comprising a nucleotide sequence having at least 85% identity to SEQ ID NO:

7.

5. The nucleotide sequence of claim 4 wherein said nucleotide sequence is isolated from Sus genus.

6. The nucleotide sequence of claim 5 wherein said Sus genus is Sus scrofa.

7. A method of producing porcine intrinsic factor comprising the steps of:

(a) isolating a nucleic acid comprising a nucleotide sequence;

i) encoding porcine intrinsic factor comprising an amino acid sequence having at least 85% identity to SEQ ID NO;

9, or

ii) having at least 85% identity to SEQ ID NO;

7;

(b) constructing a vector comprising;

i) said isolated nucleic acid operably linked to ii) a regulatory sequence; and

(c) introducing said vector into a host cell for a time and under conditions sufficient for expression of said porcine intrinsic factor.

8. A vector comprising:

(a) an isolated nucleic acid comprising a nucleotide sequence;

i) encoding a polypeptide comprising an amino acid sequence having at least 85% identity to SEQ ID NO;

9, or

ii) having at least 85% identity to SEQ ID NO;

7, operably linked to

(b) a regulatory sequence.

9. An isolated nucleic acid complementary to a nucleotide sequence encoding porcine intrinsic factor, wherein the amino acid sequence of said porcine intrinsic factor has at least 85% identity to SEQ ID NO:

9.

10. An isolated nucleic acid sequence complementary to a nucleotide sequence having at least 85% identity to SEQ ID NO:

7.

11. A method of producing porcine intrinsic factor comprising the steps of:

(a) isolating a nucleic acid complementary to a nucleotide sequence;

i) encoding porcine intrinsic factor comprising an amino acid sequence having at least 85% identity to SEQ ID NO;

9, or ii) having at least 85% identity to SEQ ID NO;

7;

(b) constructing a vector comprising;

i) said isolated nucleic acid operably linked to ii) a regulatory sequence; and

(c) introducing said vector into a host cell for a time and under conditions sufficient for expression of said porcine intrinsic factor.

12. A vector comprising:

(a) an isolated nucleic acid complementary to a nucleotide sequence;

i) encoding a polypeptide comprising an amino acid sequence having at least 85% identity to SEQ ID NO;

9, or ii) having at least 85% identity to SEQ ID NO;

7, operably linked to (b) a regulatory sequence.