Multi-species polynucleotide control sequences

1. An expression cassette comprising a gene-of-interest and a polynucleotide control sequence which directs the expression of polypeptides in at least three of the following groups of host cells: algae, fungi, yeasts, gram-positive bacteria, gram-negative bacteria, wherein the polynucleotide control sequence comprises a polynucleotide sequence of the general formula T-T-G-A-C—W-N(o)-Y 1 -A-Y 2 -A-A-T-H 1 -H 2 -N(p)-S 1 -S 2 -W-K 1 -K 2 -N(q)-H 3 -M 1 -M 2 -H 4 -A-T-G, (formula I) wherein the polynucleotide sequence is selected from any one of SEQ ID NOS: 1 to 51, wherein

N is any of the nucleotides A, C, G and T, and N(o) is 16, 17 or 18 nucleotides long;

N(p) is 22 nucleotides long;

N(q) is 2, 3, 4, 5 or 6 nucleotides long; and

each individual N in N(o), N(p), and N(q) are the same or different;

W is any of the nucleotides A and T;

Y is any of the nucleotides C and T, and Y 1 and Y 2 are the same or different;

H is any of the nucleotides A, C, and T and H 1 , H 2 , H 3 and H 4 are the same or different;

S is any of the nucleotides G and C, and S 1 and S 2 are the same or different;

K is any of the nucleotides G and T, and K 1 and K 2 are the same or different;

M is any of the nucleotides A and C, and M 1 and M 2 are the same or different;

the AG-content of N(o) is below 50%;

the A-content of N(o) is below 35%;

the AG-content within each stretch of 6 nucleotides in N(o) is below 66%;

the AG-content of N(p) is below 60%;

the AG-content within each stretch of 6 nucleotides in N(p) is below 83%;

N(o) and N(p) do not contain two consecutive G's; and

the G-content of N(q) is below 50%,

wherein the polynucleotide control sequence is either heterologous to the host cell or to the gene-of-interest coding for the polypeptide.

2. The expression cassette according to claim 1 , wherein N(o) is 17 nucleotides long; N(p) is 22 nucleotides long; and N(q) is 3 or 4 nucleotides long.

3. The expression cassette according to claim 1 , wherein the polynucleotide control sequence is selected from the group consisting of SEQ ID NO. 46 to 51.

4. The expression cassette according to claim 1 which directs the expression of polypeptides in at least four of the following genera: Escherichia, Streptomyces, Bacillus, Gluconobacter, Pseudomonas, Clostridium, Saccharomyces, Kluyveromyces, Pichia, Penicillium, Aspergillus, Mortierella, Chrysosporium, Acremonium, Trichoderma.

5. A vector comprising an expression cassette, wherein the expression cassette comprises a polynucleotide sequence and a gene-of-interest, wherein the polynucleotide control sequence comprises a polynucleotide sequence of the general formula

T-T-G-A-C-W-N(o)-Y 1 -A-Y 2 -A-A-T-H 1 -H 2 -N(p)-S 1 -S 2 -W-K 1 -K 2 -N(q)-H 3 -M 1 -M 2 -H 4 -A-T-G, (formula I) wherein the polynucleotide sequence is selected from any one of SEQ ID NOS: 1 to 51, wherein

N is any of the nucleotides A, C, G and T, and N(o) is 16, 17 or 18 nucleotides long;

N(p) is 22 nucleotides long;

N(q) is 2, 3, 4, 5 or 6 nucleotides long; and

each individual N in N(o), N(p), and N(q) are the same or different;

W is any of the nucleotides A and T;

Y is any of the nucleotides C and T, and Y 1 and Y 2 are the same or different;

H is any of the nucleotides A, C, and T and H 1 , H 2 , H 3 and H 4 are the same or different;

S is any of the nucleotides G and C, and S 1 and S 2 are the same or different;

K is any of the nucleotides G and T, and K 1 and K 2 are the same or different;

M is any of the nucleotides A and C, and M 1 and M 2 are the same or different;

the AG-content of N(o) is below 50%;

the A-content of N(o) is below 35%;

the AG-content within each stretch of 6 nucleotides in N(o) is below 66%;

the AG-content of N(p) is below 60%;

the AG-content within each stretch of 6 nucleotides in N(p) is below 83%;

N(o) and N(p) do not contain two consecutive G's; and

the G-content of N(q) is below 50%,

wherein the polynucleotide control sequence directs expression of the gene-of-interest in at least three of the following groups of host cells: algae, fungi, yeast, gram-positive bacteria, gram-negative bacteria, wherein the polynucleotide control sequence is either heterologous to the host cell or to the gene-of-interest coding for the polypeptide.

6. A host cell comprising the vector according to claim 5 .

7. A method of producing a polypeptide, a primary or secondary metabolite, an antibody or a pharmaceutical with a host cell according to claim 6 .

8. A method according to claim 7 , wherein the host cell is a cloning host, and wherein the polypeptide, primary or secondary metabolite, antibody or pharmaceutical is produced in a production host, which method comprises using the same polynucleotide control sequence in both cloning host and production host.

9. The method according to claim 8 , wherein the polynucleotide control sequence is a control sequence.

10. The method according to claim 8 , wherein the production host is of a different species than the cloning host.

11. A method for cloning a polynucleotide sequence comprising:

(a) cloning a polynucleotide sequence in front of a selectable marker gene on a vector according to the invention;

(b) transfecting a first species with the vector from step (a);

(c) obtaining clones after selection for active transcription of the selectable marker gene;

(d) isolating DNA from these clones;

(e) transfecting another species with the isolated DNA,

wherein steps (c) to (e) are repeated minimally 2 times;

wherein the polynucleotide sequence is of the general formula

T-T-G-A-C-W-N(o)-Y 1 -A-Y 2 -A-A-T-H 1 -H 2 -N(p)-S 1 -S 2 -W-K 1 -K 2 -N(q)-H 3 -M 1 -M 2 -H 4 -A-T-G, (formula I) wherein the polynucleotide sequence is selected from any one of SEQ ID NOS: 1 to 51, wherein

N is any of the nucleotides A, C, G and T, and N(o) is 16, 17 or 18 nucleotides long;

N(p) is 22 nucleotides long;

N(q) is 2, 3, 4, 5 or 6 nucleotides long; and

each individual N in N(o), N(p), and N(q) are the same or different;

W is any of the nucleotides A and T;

Y is any of the nucleotides C and T, and Y 1 and Y 2 are the same or different;

H is any of the nucleotides A, C, and T and H 1 , H 2 , H 3 and H 4 are the same or different;

S is any of the nucleotides G and C, and S 1 and S 2 are the same or different;

K is any of the nucleotides G and T, and K 1 and K 2 are the same or different;

M is any of the nucleotides A and C, and M 1 and M 2 are the same or different;

the AG-content of N(o) is below 50%;

the A-content of N(o) is below 35%;

the AG-content within each stretch of 6 nucleotides in N(o) is below 66%;

the AG-content of N(p) is below 60%;

the AG-content within each stretch of 6 nucleotides in N(p) is below 83%;

N(o) and N(p) do not contain two consecutive G's; and

the G-content of N(q) is below 50%.

12. A molecular biology kit comprising at least one reagent and a polynucleotide control sequence according to claim 1 .

13. The expression cassette according to claim 1 , wherein the polynucleotide control sequence is selected from the group consisting of SEQ ID NO. 46.

14. The expression cassette according to claim 1 , wherein the polynucleotide control sequence is selected from the group consisting of SEQ ID NO. 47.

15. The expression cassette according to claim 1 , wherein the polynucleotide control sequence is selected from the group consisting of SEQ ID NO. 48.

16. The expression cassette according to claim 1 , wherein the polynucleotide control sequence is selected from the group consisting of SEQ ID NO. 49.

17. The expression cassette according to claim 1 , wherein the polynucleotide control sequence is selected from the group consisting of SEQ ID NO. 50.

18. The expression cassette according to claim 1 , wherein the polynucleotide control sequence is selected from the group consisting of SEQ ID NO. 51.