Table 1 Bacterial strains, plasmids and oligonucleotides used in this study

Escherichia coli

recA1, endA1, gyrA96, thi-1, hsdR17 (r_k^-, m_k⁺),

[16]

XL1-Blue

supE44, relA1, -, lac [F', proAB, lacI^q, lacZΔM15, Tn10(Tc^r)]

Cupriavidus necator H16

Wildtype (PHA⁺)

DSMZ 428

Plasmids

pCRII

Amp, ColE1 origin

Initrogen

pBHR68

PHB biosynthesis operon from C. necator in pBluescriptSK^-

[17]

pHAS

pET-14b containing NdeI/BamHI inserted phaC gene from C. necator

[18]

pUC57-MOG

DNA fragment encoding N terminal 1–173 amino acids of MOG from mouse inserted in SmaI site of pUC57

This study

pUC57-IL2

DNA fragment encoding N terminal 60–169 amino acids of IL2 from mouse inserted in SmaI site of pUC57

This study

pCR-phaP

pCRII containing phaP gene from C. necator

This study

pHAS-phaP

pHAS containing the phaP gene inserted into XbaI/NdeI sites

This study

pHAS-phaP-MOG

pHAS-phaP containing the MOG encoding DNA fragment inserted into NdeI/BamHI sites

This study

pHAS-phaP-MOG

pHAS-phaP containing the IL2 encoding DNA fragment inserted into NdeI/BamHI sites

This study

pUC57-phaP-MOG

DNA fragment encoding PhaP inserted into XbaI and NdeI site of plasmid pUC57-MOG

This study

pUC57-phaP-IL2

DNA fragment encoding PhaP inserted into XbaI and NdeI site of plasmid pUC57-IL2

This study

pBHR68-phaP-MOG

DNA fragment encoding PhaP-MOG fusion protein subcloned from pUC57-phaP-MOG via XbaI and BamHI into pBHR68

This study

pBHR68-phaP-IL2

DNA fragment encoding PhaP-IL2 fusion protein subcloned from pUC57-phaP-IL2 via XbaI and BamHI into pBHR68

This study

Oligonucleotides

phaP-XbaI-NdeI

5'-aaaaatctagaaaaaggagatatacatatgatcctc-accccggaacaag-3'

This study

phaP-XbaI

5'-aaaaatctagaaaaaggagatatacgtatgatcctc-accccggaacaag-3'

This study

phaP-NdeI

5'-aaaaacatatggtggtgatggtgatgcgagccgcgt-ttatcatcatcatcggcagccgtcgtcttc-3'

This study