Recombinant Lactococcus lactis NZ9000 secretes a bioactive kisspeptin that inhibits proliferation and migration of human colon carcinoma HT-29 cells

Table 2 Primers used in this study

Primer	Sequence (5′ → 3′)^a	Description and location
Primer 1	ATCCATGGGTATGAAAAAAAAGATTAT	Upstream of SPP2-F, NcoI^b
Primer 2	CATGCATGCTGCGTCACAAGTCGACGATA	Downstream of SPP2-R, SphI
Primer 3	CATGCATGCATGAACTCACTGGTTTCTT	Upstream of kiss1-F, SphI^c
Primer 4	CCCAAGCTTCAGCCCCGCCCAGCGCTTC	Downstream of kiss1-R, HindIII
Primer 5	ACCACAGTCCATGCCATCAC	Upstream of GAPDH for RT-PCR
Primer 6	TCCACCACCCTGTTGCTGTA	Downstream of GAPDH for RT-PCR
Primer 7	GGT GGA CCG GAT GTT CCC	Upstream of MMP-9 for RT-PCR
Primer 8	GCC CAC CTC TCC TCC	Downstream of MMP-9 for RT-PCR

^aThe added restriction enzyme sites are underlined
^bSPP2 includes Usp45 signal peptide and LEISSTCDA synthetic propeptide
^c Homo sapiens KiSS-1 metastasis suppressor: gb:BC022819.1 (cDNA clone MGC:39258 IMAGE:5443510)

ISSN: 1475-2859