TABLE 3.

cDNA amplification and vector sequencing primers for fruit bat and vesper bat tetherins^a

Tetherin source and type	cDNA amplification primer	Primer sequence (5′ > 3′)	Direction	Target region	Successful combinations	PCR TA (°C)
Pteropus alecto
Tetherin	PaF1_JH_119	TCACTGCAAGGGGTTCTCTC	Forward	5′ UTR	119/121−2	60
	PaF2_JH_120	GGAAACTTCACTGCAAGGGG	Forward	5′ UTR	120/121−2	60
	PaR1_JH_121	CTTCTCCCAGCTTTGTTGCC	Reverse	3′ UTR
	PaR2_JH_122	CTCCTCTCCCCCAAAATGTC	Reverse	3′ UTR
Miniopterus schreibersii
Tetherin	MiniF1_JH_466	CCCACAAACTCCCTACACCC	Forward	5′ UTR	466/468	60
	MiniF2_JH_467	ATGCTAATGAAGGGGCGGGG	Forward	5′ UTR	466/469	62
	MiniR1_JH_468	CTGTCTGTCTTCCTGGGAC	Reverse	3′ UTR	467/469	62
	MiniR2_JH_469	GGACAGGTCAGGGAAACCAA	Reverse	3′ UTR
Myotis macropus and Myotis ricketti
Tetherins A, C, and D	MyoF1_JH_479	TCCACTGCATCCCTCTG	Forward	5′ UTR	479/481−4	60
	MyoF2_JH_480	ATGGCACCCACTTTTTAC	Forward	5′ UTR	480/481−4	60
	MyoR1_JH_481	TCAGCCAGGTTAGAATGTG	Reverse	3′ UTR
	MyoR2_JH_482	TCCTTGGGCAAACAGCTCTC	Reverse	3′ UTR
	MyoR3_JH_483	CAGGAAACTCTCAGAAAAG	Reverse	3′ UTR
	MyoR4_JH_484	CATCTTTCCAAGACCACA	Reverse	3′ UTR
Tetherins B and E	MyoF3_JH_474	GCTCCTGTGCATCCCTCTGG	Forward	5′ UTR	474/478	60
	MyoR5_JH_478	CCTGGTTAGAATGTGCTTT	Reverse	3′ UTR

^aPrimers used to amplify tetherin homologues from cDNA generated from Myotis macropus, M. ricketti, and Miniopterus schreibersii. Combinations of forward and reverse primers identified as capable of amplifying tetherin are indicated alongside the highest annealing temperature at which amplification was successful. UTR, untranslated region; T_A, annealing temperature.