Supplement data
Table 1 Oligonucleotide primers for amplification of R. affinis and S. kuhlii TTR cDNAs
and intron1 fragments
Primer
Nucleotide sequence (53)
TTR_degen_#1F
Direction
CGACACCAAGTGCCCTCTGATGGTIAARRT
Forward*
TTR_degen_#1R
GGTGAACACCACCTCGGCGTACTCRTGRAAIGG
Reverse*
5GSP1_BAT
AGTTGTGAGCCCGTGGAGCTCTCCAAATTCACTGGTTTTCC
Reverse
5GSP3_BAT
TGTGGCGAACGGCTCCCAGGTCTCATC
Reverse
5GSP2_BAT
TGATGAGACCTGGGAGCCGTTCGCCACAG
Forward
3inter_bat1
TACTGGAAGGAGCTGGGCATCTCGCCCTTTCACGAGTACG
Reverse
BAT_fullseqF
AGAGGCCCACTGATTCTGGGCAGGATGGCTTCTCGG
Forward
BAT_fullseqR
TCATTCTTTGGGGGCGCTGACGAGGGCTGTGGTGGAG
Reverse
SKbatfull1_F
TGCCGGGATGGCTGCCCTCCGTCTCCTGCTCC
Forward
3_SKTTRFull2
TCCCTGCCTGGGCTGCTAACATAGCTTATGAGGC
Reverse
SKbat_intron1F
TGGCTGCCCTCCGTCTCCTGCTCCTCTG
Forward
SKbat_intron1R
TCGGCAGTCTTCTTGAACACCTTCACGGCCACG
Reverse
* the sequences were from Manzon et al. 2007.
I = deoxyinosine
Figure legend of supplement data
Fig. 1
Determination of T4 binding proteins in bat serum.
Purified human TTR (1) and albumin (2), and aliquots (5 l) of serum from
S. kuhlii (3) and human (HS) were incubated with 2 g of T4-FITC for 5 min.
The proteins were separated by native-PAGE (10% resolving gel), followed by
staining with Coomassie blue R-250 (Coomassie). Bound T4 was visualized
under UV (Fluorescence). Positions of thyroxine binding globulin (TBG),
albumin (ALB) and TTR (TTR) in human serum were indicated. Closed arrow
head, opened arrow head, and arrow indicate T4-FITC that bound to TBG,
albumin, and TTR of S. kuhlii, respectively.
Fig. 2
Nucleotide sequence and derived amino acid sequence of TTR cDNA from
R. affinis.
a: Strategy to obtain the full-length sequence of TTR cDNA. R. affinis TTR
cDNA is represented as a stripe bar. Arrows indicate the TTR cDNA fragment
generated by degenerated RT-PCR (nucleotide 96-367), the 5RACE
(nucleotide 1-190) and the 3RACE (nucleotide 218-618).
b: Nucleotide numbers are shown on the right. The amino acids in the signal
peptide sequence are numbered -20 to -1. The N-terminal residue of the mature
TTR is designated as +1. The first inframe stop codon is represented by 3
asterisks. The putative polyadenylation signal in the 3-untranslated region of
cDNA is underlined.
Fig. 3
Nucleotide sequence and derived amino acid sequence of TTR cDNA from
S. kuhlii.
a: Strategy to obtain the full-length sequence of TTR cDNA from R. affinis.
Dot bar, S. kuhlli TTR cDNA; arrows, the TTR cDNA fragments generated by
RT-PCR (132-340), 5RACE (1-220), and 3RACE (317-609).
b: The nucleotide numbers are shown on the right, and numbering of amino
acids is as described in Fig. 2.
Fig. 4
The nucleotide sequences of the TTR intron 1 flanking the continuous (a) and
discontinuous (b) base deletion characteristics of the extant Chiroptera. The
nucleotide numbers were on the right of the sequence.